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Статті в журналах з теми "Human proteases"
1
Sampson, M. T., and A. K. Kakkar. "Coagulation proteases and human cancer." Biochemical Society Transactions 30, no. 2 (April 1, 2002): 201–7. http://dx.doi.org/10.1042/bst0300201.
Повний текст джерелаАнотація:
Tumours are capable of activating blood coagulation through the expression of procoagulant molecules such as tissue factor, cancer procoagulant and hepsin. Initiation of the clotting cascade results in the generation of the activated serine proteases factor VIIa, factor Xa and thrombin. These proteases act via protease-activated receptors and tissue factor to alter gene expression, thereby modulating tumour cell growth, invasion, metastasis and angiogenesis.
2
Weksler, BB, EA Jaffe, MS Brower, and OF Cole. "Human leukocyte cathepsin G and elastase specifically suppress thrombin- induced prostacyclin production in human endothelial cells." Blood 74, no. 5 (October 1, 1989): 1627–34. http://dx.doi.org/10.1182/blood.v74.5.1627.1627.
Повний текст джерелаАнотація:
Abstract Polymorphonuclear leukocytes (PMN) when activated release products that can potentially injure endothelial cells or alter endothelial function. Exposure of cultured human umbilical vein endothelial cells to cathepsin G and elastase isolated from human PMN at concentrations reached in vivo (100 ng/mL to 10 micrograms/mL) selectively inhibited thrombin-induced prostacyclin production and the thrombin-induced rise in cytosolic free calcium ([Ca++]i) concentration. These proteases also blocked thrombin-induced release of arachidonic acid from prelabeled endothelial cells (EC). In contrast, induction of prostacyclin (PGI2) production by arachidonate, histamine, or the calcium ionophore A23187 was not altered by treatment of EC with these proteases. The effects of the proteases were concentration-dependent, were blocked by serum or serum protease inhibitors, and were reversed when the endothelial cells were further cultured for 24 hours in the absence of the proteases. Elastase, but not cathepsin G, also produced detachment of endothelial cells. Thus, the major leukocyte proteases selectively suppress thrombin-induced prostacyclin production by human vascular endothelial cells and may alter the hemostatic balance at sites of PMN activation.
3
Weksler, BB, EA Jaffe, MS Brower, and OF Cole. "Human leukocyte cathepsin G and elastase specifically suppress thrombin- induced prostacyclin production in human endothelial cells." Blood 74, no. 5 (October 1, 1989): 1627–34. http://dx.doi.org/10.1182/blood.v74.5.1627.bloodjournal7451627.
Повний текст джерелаАнотація:
Polymorphonuclear leukocytes (PMN) when activated release products that can potentially injure endothelial cells or alter endothelial function. Exposure of cultured human umbilical vein endothelial cells to cathepsin G and elastase isolated from human PMN at concentrations reached in vivo (100 ng/mL to 10 micrograms/mL) selectively inhibited thrombin-induced prostacyclin production and the thrombin-induced rise in cytosolic free calcium ([Ca++]i) concentration. These proteases also blocked thrombin-induced release of arachidonic acid from prelabeled endothelial cells (EC). In contrast, induction of prostacyclin (PGI2) production by arachidonate, histamine, or the calcium ionophore A23187 was not altered by treatment of EC with these proteases. The effects of the proteases were concentration-dependent, were blocked by serum or serum protease inhibitors, and were reversed when the endothelial cells were further cultured for 24 hours in the absence of the proteases. Elastase, but not cathepsin G, also produced detachment of endothelial cells. Thus, the major leukocyte proteases selectively suppress thrombin-induced prostacyclin production by human vascular endothelial cells and may alter the hemostatic balance at sites of PMN activation.
4
Avidano, Michael A., Cheryl S. Cotter, Scott P. Stringer, and Gregory S. Schultz. "Analysis of protease activity in human otitis media." Otolaryngology–Head and Neck Surgery 119, no. 4 (October 1998): 346–51. http://dx.doi.org/10.1016/s0194-5998(98)70076-2.
Повний текст джерелаАнотація:
Cronic otitis media is a common problem associated with a nonintact tympanic membrane frequently involving Staphylococcus aureus and Pseudomonas aeruginosa. The virulence of Pseudomonas bacteria is related to the production of two matrix metalloproteinases, elastase and alkaline protease. Serine proteases, such as neutrophil elastase, are produced by the host inflammatory response. These proteases are thought to contribute to tissue destruction and assist bacterial invasion during infection. This preliminary study was done to identify protease activity in otorrhea samples from patients with otitis media and a nonintact tympanic membrane and to examine the ability of selective protease inhibitors to decrease protease activity. Ilomostat (galardin) is a synthetic, specific inhibitor of matrix metalloproteinases including P. aeruginosa elastase and alkaline protease, where-as α1-antitrypsin inhibits serine proteases including neutrophil elastase. Samples were collected and cultured from 20 patients with otorrhea resulting from tympanic membrane perforations or pressure-equalization tubes. A protease assay that used azocasein as the substrate was used to quantify protease activity, with and without addition of selective protease inhibitors. Cultures revealed P. aeruginosa alone in 7 samples, P. aeruginosa plus other organisms in 10, and S. aureus alone in 3. Protease activity was detected in 15 (75%) of the samples. A statistically significant ( p < 0.05) decrease in protease activity was seen with the addition of α1-antitrypsin or Ilomostat plus α1-antitrypsin, but not with Ilomostat alone. Analyzing the 10 samples with the highest protease activity, a statistically significant decrease in activity was seen with Ilomostat or αα1-antitrypsin alone and with both Ilomostat and α1-antitrypsin together. Bacteriologic type, source of sample, age and gender of the subject, and duration of infection were not significantly related to protease activity. This is the first study to quantify protease activity and inhibition by selective protease inhibitors in human otorrhea. Protease inhibitors effectively decrease protease activity in most cases and in addition to standard antibiotic therapy might prove beneficial in the treatment of otitis media with a nonintact tympanic membrane. This study supports future clinical investigations into the role of proteases and inhibition of protease activity in the treatment of otitis media.
5
Duvezin-Caubet, Stéphane, Mirko Koppen, Johannes Wagener, Michael Zick, Lars Israel, Andrea Bernacchia, Ravi Jagasia, et al. "OPA1 Processing Reconstituted in Yeast Depends on the Subunit Composition of the m-AAA Protease in Mitochondria." Molecular Biology of the Cell 18, no. 9 (September 2007): 3582–90. http://dx.doi.org/10.1091/mbc.e07-02-0164.
Повний текст джерелаАнотація:
The morphology of mitochondria in mammalian cells is regulated by proteolytic cleavage of OPA1, a dynamin-like GTPase of the mitochondrial inner membrane. The mitochondrial rhomboid protease PARL, and paraplegin, a subunit of the ATP-dependent m-AAA protease, were proposed to be involved in this process. Here, we characterized individual OPA1 isoforms by mass spectrometry, and we reconstituted their processing in yeast to identify proteases involved in OPA1 cleavage. The yeast homologue of OPA1, Mgm1, was processed both by PARL and its yeast homologue Pcp1. Neither of these rhomboid proteases cleaved OPA1. The formation of small OPA1 isoforms was impaired in yeast cells lacking the m-AAA protease subunits Yta10 and Yta12 and was restored upon expression of murine or human m-AAA proteases. OPA1 processing depended on the subunit composition of mammalian m-AAA proteases. Homo-oligomeric m-AAA protease complexes composed of murine Afg3l1, Afg3l2, or human AFG3L2 subunits cleaved OPA1 with higher efficiency than paraplegin-containing m-AAA proteases. OPA1 processing proceeded normally in murine cell lines lacking paraplegin or PARL. Our results provide evidence for different substrate specificities of m-AAA proteases composed of different subunits and reveal a striking evolutionary switch of proteases involved in the proteolytic processing of dynamin-like GTPases in mitochondria.
6
Menou, Awen, JanWillem Duitman, Pauline Flajolet, Jean-Michel Sallenave, Arnaud André Mailleux, and Bruno Crestani. "Human airway trypsin-like protease, a serine protease involved in respiratory diseases." American Journal of Physiology-Lung Cellular and Molecular Physiology 312, no. 5 (May 1, 2017): L657—L668. http://dx.doi.org/10.1152/ajplung.00509.2016.
Повний текст джерелаАнотація:
More than 2% of all human genes are coding for a complex system of more than 700 proteases and protease inhibitors. Among them, serine proteases play extraordinary, diverse functions in different physiological and pathological processes. The human airway trypsin-like protease (HAT), also referred to as TMPRSS11D and serine 11D, belongs to the emerging family of cell surface proteolytic enzymes, the type II transmembrane serine proteases (TTSPs). Through the cleavage of its four major identified substrates, HAT triggers specific responses, notably in epithelial cells, within the pericellular and extracellular environment, including notably inflammatory cytokine production, inflammatory cell recruitment, or anticoagulant processes. This review summarizes the potential role of this recently described protease in mediating cell surface proteolytic events, to highlight the structural features, proteolytic activity, and regulation, including the expression profile of HAT, and discuss its possible roles in respiratory physiology and disease.
7
Puente, X. S., L. M. Sánchez, A. Gutiérrez-Fernández, G. Velasco, and C. López-Otín. "A genomic view of the complexity of mammalian proteolytic systems." Biochemical Society Transactions 33, no. 2 (April 1, 2005): 331–34. http://dx.doi.org/10.1042/bst0330331.
Повний текст джерелаАнотація:
Proteolytic enzymes play an essential role in different physiological processes, including development, reproduction and host defence, as well as in numerous pathologies, like inflammatory diseases, neurological disorders or cancer. The completion of the human genome sequence allowed us to determine that more than 2% of all human genes are proteases or protease inhibitors, reflecting the importance of proteolysis in human biology. To understand better the complexity of proteases in human and other model organisms, we have used the available genome sequences of different mammalian organisms, including mouse, rat and chimpanzee, to identify and compare their degradomes, the complete set of protease genes in these species. Surprisingly, the rodent protease complement is more complex when compared with that of primates, mainly due to the expansion of protease families implicated in reproduction and host defence. Similarly, most differences between human and chimpanzee proteases are found in genes implicated in the immune system, which might explain some of the differences between both organisms. We have also found several genes implicated in reproduction, nutrition and the immune system, which are functional in rat, mouse or chimpanzee, but have been inactivated by mutations in the human lineage. These findings suggest that pseudogenization of specific protease genes has been a mechanism contributing to the evolution of the human genome. Finally, we found that proteases implicated in human hereditary diseases, and especially in neurodegenerative disorders, are highly conserved among mammals.
8
Szabo, Roman, Qingyu Wu, Robert Dickson, Sarah Netzel-Arnett, Toni Antalis, and Thomas Bugge. "Type II transmembrane serine proteases." Thrombosis and Haemostasis 90, no. 08 (2003): 185–93. http://dx.doi.org/10.1160/th03-02-0071.
Повний текст джерелаАнотація:
SummaryThe recent availability of human and mouse genome sequences and expressed sequence tag databases facilitated the identification of a large new family of membrane anchored serine proteases, the type II transmembrane serine proteases or TTSPs. Analyses of human inherited disorders and gene targeting studies in mice have revealed that several members of this new protease family have critical functions in development and health. Preliminary studies also suggest that aberrant expression of type II transmembrane serine proteases may be linked to disease progression. The knowledge gathered thus far of the genetics, physiology, and pathology of this interesting new serine protease family will be reviewed here in brief.
9
Aimes, Ronald, Andries Zijlstra, John Hooper, Steven Ogbourne, Mae-Le Sit, Simone Fuchs, David Gotley, James Quigley, and Toni Antalis. "Endothelial cell serine proteases expressed during vascular morphogenesis and angiogenesis." Thrombosis and Haemostasis 89, no. 03 (2003): 561–72. http://dx.doi.org/10.1055/s-0037-1613388.
Повний текст джерелаАнотація:
SummaryMany serine proteases play important regulatory roles in complex biological systems, but only a few have been linked directly with capillary morphogenesis and angiogenesis. Here we provide evidence that serine protease activities, independent of the plasminogen activation cascade, are required for microvascular endothelial cell reorganization and capillary morphogenesis in vitro. A homology cloning approach targeting conserved motifs present in all serine proteases, was used to identify candidate serine proteases involved in these processes, and revealed 5 genes (acrosin, testisin, neurosin, PSP and neurotrypsin), none of which had been associated previously with expression in endothelial cells. A subsequent gene-specific RT-PCR screen for 22 serine proteases confirmed expression of these 5 genes and identified 7 additional serine protease genes expressed by human endothelial cells, urokinase-type plasminogen activator, protein C, TMPRSS2, hepsin, matriptase/ MT-SP1, dipeptidylpeptidase IV, and seprase. Differences in serine protease gene expression between microvascular and human umbilical vein endothelial cells (HUVECs) were identified and several serine protease genes were found to be regulated by the nature of the substratum, ie. artificial basement membrane or fibrillar type I collagen. mRNA transcripts of several serine protease genes were associated with blood vessels in vivo by in situ hybridization of human tissue specimens. These data suggest a potential role for serine proteases, not previously associated with endothelium, in vascular function and angiogenesis.
10
Ehrhardt, Katrin, Natalie Steck, Reinhild Kappelhoff, Stephanie Stein, Florian Rieder, Ilyssa O. Gordon, Erin C. Boyle, et al. "Persistent Salmonella enterica Serovar Typhimurium Infection Induces Protease Expression During Intestinal Fibrosis." Inflammatory Bowel Diseases 25, no. 10 (May 9, 2019): 1629–43. http://dx.doi.org/10.1093/ibd/izz070.
Повний текст джерелаАнотація:
AbstractBackgroundIntestinal fibrosis is a common and serious complication of Crohn’s disease characterized by the accumulation of fibroblasts, deposition of extracellular matrix, and formation of scar tissue. Although many factors including cytokines and proteases contribute to the development of intestinal fibrosis, the initiating mechanisms and the complex interplay between these factors remain unclear.MethodsChronic infection of mice with Salmonella enterica serovar Typhimurium was used to induce intestinal fibrosis. A murine protease-specific CLIP-CHIP microarray analysis was employed to assess regulation of proteases and protease inhibitors. To confirm up- or downregulation during fibrosis, we performed quantitative real-time polymerase chain reaction (PCR) and immunohistochemical stainings in mouse tissue and tissue from patients with inflammatory bowel disease. In vitro infections were used to demonstrate a direct effect of bacterial infection in the regulation of proteases.ResultsMice develop severe and persistent intestinal fibrosis upon chronic infection with Salmonella enterica serovar Typhimurium, mimicking the pathology of human disease. Microarray analyses revealed 56 up- and 40 downregulated proteases and protease inhibitors in fibrotic cecal tissue. Various matrix metalloproteases, serine proteases, cysteine proteases, and protease inhibitors were regulated in the fibrotic tissue, 22 of which were confirmed by quantitative real-time PCR. Proteases demonstrated site-specific staining patterns in intestinal fibrotic tissue from mice and in tissue from human inflammatory bowel disease patients. Finally, we show in vitro that Salmonella infection directly induces protease expression in macrophages and epithelial cells but not in fibroblasts.ConclusionsIn summary, we show that chronic Salmonella infection regulates proteases and protease inhibitors during tissue fibrosis in vivo and in vitro, and therefore this model is well suited to investigating the role of proteases in intestinal fibrosis.
Дисертації з теми "Human proteases"
1
Lourbakos, Afrodite 1972. "Activation of human protease-activated receptors by proteases from a periodontal pathogen." Monash University, Dept. of Biochemistry and Molecular Biology, 2001. http://arrow.monash.edu.au/hdl/1959.1/8876.
Повний текст джерела
2
Agrotis, Alexander George. "The regulation of human autophagy by ATG4 proteases." Thesis, University College London (University of London), 2018. http://discovery.ucl.ac.uk/10048926/.
Повний текст джерелаАнотація:
Autophagy is an important intracellular degradation pathway that delivers cytoplasmic material to lysosomes via double-membrane organelles called autophagosomes. Lipidation of ubiquitin- like LC3/GABARAP proteins on the autophagosome membrane is essential for autophagy. The cysteine protease ATG4 executes C-terminal processing (priming) of newly-synthesised LC3/GABARAP to enable subsequent lipidation. ATG4 is also proposed to be important for deconjugating/delipidating LC3/GABARAP from autophagosome membranes, although the ex- act purpose of this is unclear in mammals. Four ATG4 isoforms (ATG4A-D) exist in mammals, however the functional redundancy of these proteins in cells is poorly understood. The aim of this thesis is to characterise the redundancy of human ATG4 proteins and investigate their deconjugation roles in cells. In Chapter 3, I show that human HAP1 and HeLa cells lacking ATG4B exhibit a severe but incomplete defect in LC3/GABARAP processing and autophagy. By further genetic depletion of ATG4 isoforms I uncover that ATG4A, ATG4C and ATGD all contribute to residual priming activity, which is sufficient to enable lipidation of GABARAPL1 on autophagosomes. In Chapter 4, I reveal that delipidation of LC3/GABARAP by ATG4 isoforms is not essential for autophagic degradation of the cargo receptor p62/SQSTM1, arguing that delip- idation by ATG4 has limited importance in mammalian autophagy compared to LC3/GABARAP priming. Finally, I report the discovery of a novel deconjugation function of ATG4 isoforms in the removal of LC3/GABARAP conjugates from endogenous proteins such as ATG3 and ATG7. Thus, I provide the first evidence that LC3/GABARAP can act as a protein modifier and that ATG4 can reverse such modifications, opening up a new avenue of future research to under- stand the functional relevance of this phenomenon.
3
Ranjit, Najju. "Characterisation of proteases involved in proteolytic degradation of haemoglobin in the human hookworm Necator americanus." Thesis, Queensland University of Technology, 2008. https://eprints.qut.edu.au/20651/1/Najju_Ranjit_Thesis.pdf.
Повний текст джерелаАнотація:
With over a billion people infected world wide, hookworms are considered as important human pathogens, particularly in developing countries which have the highest rates of infections. Hookworms reside in the gastrointestinal tract of the host where they continuously feed on blood, leading to conditions such as chronic irondeficiency anaemia. The majority of blood-feeding parasites rely on proteins found in blood to provide many of their nutritional requirements for growth, reproduction and survival. Of the numerous proteins found in blood, haemoglobin (Hb) is one of the most abundant. In order to acquire amino acids for protein synthesis, it is thought that haematophagous parasites degrade Hb using various classes of endo- and exoproteases, in a manner similar to that which occurs in catabolism of proteins in mammalian cellular lysosomes. This study identified and characterised proteases involved in the Hb degradation process in the human hookworm, Necator americanus, in order to identify potential candidate antigens for a vaccine that interrupts blood-feeding. Red blood cells ingested by hookworms are lysed to release Hb, which is cleaved by various proteases into dipeptides or free amino acids and these are taken up through the gut membrane by amino acid transporters. Proteases expressed in the intestinal tract of hookworms are thought to play a major role in this process and would therefore make good targets for vaccine candidates aimed at interrupting blood-feeding. To identify these proteases, adult hookworms (both N. americanus and Ancylostoma caninum) were sectioned and intestinal tissue was dissected via laser microdissection microscopy. RNA extracted from the dissected tissue was used to generate gut-specific cDNA, which then was used to create plasmid libraries. Each library was subjected to shotgun sequencing, and of the 480 expressed sequence tags (ESTs) sequenced from each species, 268 and 276 contigs were assembled from the N. americanus and A. caninum libraries, respectively. Nine percent of N. americanus and 6.5% of A. caninum contigs were considered novel as no homologues were identified in any published/accessible database. The gene ontology (GO) classification system was used to categorise the contigs to predicted biological functions. Only 17% and 38% of N. americanus and A. caninum contigs, respectively, were assigned GO categories, while the rest were classified as being of unknown function. The most highly represented GO categories were molecular functions such as protein binding and catalytic activity. The most abundant transcripts encoded fatty acid binding proteins, C-type lectins and activation associated secreted proteins, indicative of the diversity of functions that occur in this complex organ. Of particular interest to this study were the contigs that encoded for cysteine and metalloproteases, expanding the list of potential N. americanus haemoglobinases. In the N. americanus cDNA library, four contigs encoding for cathepsin B cysteine proteases were identified. Three contigs from the A. caninum and one contig from the N. americanus cDNA libraries encoded for metalloproteases, including astacin-like and O-sialoglycoprotein endopeptidases, neither of which had previously been reported from adult hookworms. Apart from haemoglobinases, other mRNAs encoding potential vaccine candidate molecules were identified, including anti-clotting factors, defensins and membrane proteins. This study confirmed that the gut of hookworms encodes a diverse range of proteases, some of which are likely to be involved in Hb digestion and have the potential to be hidden (cryptic) vaccine antigens. Four cysteine proteases (Na-CP-2, -3, -4 and -5) were identified from the gut cDNA library of N. americanus. All four proteases belong to the clan CA, family C1, share homology with human cathepsin B and possess a modified occluding loop. Real-time PCR indicated that all transcripts were up-regulated in the adult stage of the hookworm parasite with high levels of mRNA expression detected in gut cDNA. All four proteases were expressed in recombinant form, but only Na-CP-3 was successfully expressed in soluble form in the yeast Pichia pastoris. Proteolytic activity for Na-CP-3 was detected on a gelatin zymogen gel, however no catalytic activity was detected against the class-specific fluorogenic peptides Z-Phe-Arg-AMC and Z-Arg-Arg-AMC. Mass spectrometry analysis of the purified protein suggested that the pro-region had not been processed in trans when the protein was secreted by yeast. Incubation of Na-CP-3 in salt buffers containing dextran sulfate resulted in autoprocessing of the pro-region as detected by Western blot and catalytic activity was detected against Z-Phe-Arg-AMC. Activated Na-CP-3 did not digest intact tetrameric human Hb. The other three cysteine proteases (Na-CP-2, -4, and -5) were expressed in insoluble form in Escherichia coli. Antibodies to all four proteins (Na- CP-2 to 5) immunolocalised to the gut region of the adult worm, supporting mRNA amplification results and strongly indicated that they might play a role in nutrient acquisition. Hb digestion in blood feeding parasites such as schistosomes and Plasmodium spp. occurs via a semi-ordered cascade of proteolysis involving numerous enzymes. In Plasmodium falciparum, at least three distinct mechanistic classes of endopeptidases have been implicated in this process, and at least two classes have been implicated in schistosomes. A similar process is thought to occur in hookworms. An aspartic protease, Na-APR-1, was expressed in P. pastoris and purified protein was shown to cleave the class-specific fluorogenic peptide 7- Methoxycoumarin-4-Acetyl-GKPILFFRLK(DNP)-D-Arg-Amide. Recombinant Na- APR-1 was able to cleave intact human Hb and completely degrade the 16 kDa monomer and 32 kDa dimer within one hour. Recombinant Na-CP-3 was not able to cleave intact Hb, but was able to further digest globin fragments that had previously been digested with Na-APR-1. A clan MA metalloprotease, Na-MEP-1, was identified in gut tissue of N. americanus and was expressed in recombinant form in Hi5 insect cells using the baculovirus expression system. Recombinant Na-MEP-1 displayed proteolytic activity when assessed by gelatin zymography, but was incapable of cleaving intact Hb. However, Na-MEP-1 did cleave globin fragments which had previously been incubated with Na-APR-1 and Na-CP-3. Hb digested with all three proteases was subjected to reverse phase HPLC and peptides were analysed using Liquid Chromatography-Mass Spectrometry (LC-MS). A total of 74 cleavage sites were identified within Hb ƒ¿ and ƒÀ chains. Na-APR-1 was responsible for cleavage of Hb at the hinge region, probably unravelling the molecule so that Na- CP-3 and Na-MEP-1 could gain access to globin peptides. All three proteases were promiscuous in their subsite specificities, but the most common P1-P1�Œ residues were hydrophobic and/or bulky in nature, such as Phe, Leu and Ala. Antibodies to all three proteins (Na-APR-1, -CP-3, -MEP-1) immunolocalised to the gut region of the worm, further supporting their roles in Hb degradation. These results suggest that Hb degradation in N. americanus follows a similar pattern to that which has been described in Plasomdium falciparum. Studies conducted in this project have identified a number of potential haemoglobinases and have demonstrated that the gut region of the hookworm contains a multitude of proteases which could be targeted for production of new chemotherapies or as vaccine candidates. Results presented here also suggest that the Hb degradation process occurs in an ordered cascade, similar to those which have been reported in other haematophagous parasites. More importantly, it has been confirmed that Na-APR-1 plays a crucial role in the initiation of the Hb degradation process and therefore targeting this molecule as a vaccine candidate could provide high levels of protection against hookworm infection.
4
Ranjit, Najju. "Characterisation of proteases involved in proteolytic degradation of haemoglobin in the human hookworm Necator americanus." Queensland University of Technology, 2008. http://eprints.qut.edu.au/20651/.
Повний текст джерелаАнотація:
With over a billion people infected world wide, hookworms are considered as important human pathogens, particularly in developing countries which have the highest rates of infections. Hookworms reside in the gastrointestinal tract of the host where they continuously feed on blood, leading to conditions such as chronic irondeficiency anaemia. The majority of blood-feeding parasites rely on proteins found in blood to provide many of their nutritional requirements for growth, reproduction and survival. Of the numerous proteins found in blood, haemoglobin (Hb) is one of the most abundant. In order to acquire amino acids for protein synthesis, it is thought that haematophagous parasites degrade Hb using various classes of endo- and exoproteases, in a manner similar to that which occurs in catabolism of proteins in mammalian cellular lysosomes. This study identified and characterised proteases involved in the Hb degradation process in the human hookworm, Necator americanus, in order to identify potential candidate antigens for a vaccine that interrupts blood-feeding. Red blood cells ingested by hookworms are lysed to release Hb, which is cleaved by various proteases into dipeptides or free amino acids and these are taken up through the gut membrane by amino acid transporters. Proteases expressed in the intestinal tract of hookworms are thought to play a major role in this process and would therefore make good targets for vaccine candidates aimed at interrupting blood-feeding. To identify these proteases, adult hookworms (both N. americanus and Ancylostoma caninum) were sectioned and intestinal tissue was dissected via laser microdissection microscopy. RNA extracted from the dissected tissue was used to generate gut-specific cDNA, which then was used to create plasmid libraries. Each library was subjected to shotgun sequencing, and of the 480 expressed sequence tags (ESTs) sequenced from each species, 268 and 276 contigs were assembled from the N. americanus and A. caninum libraries, respectively. Nine percent of N. americanus and 6.5% of A. caninum contigs were considered novel as no homologues were identified in any published/accessible database. The gene ontology (GO) classification system was used to categorise the contigs to predicted biological functions. Only 17% and 38% of N. americanus and A. caninum contigs, respectively, were assigned GO categories, while the rest were classified as being of unknown function. The most highly represented GO categories were molecular functions such as protein binding and catalytic activity. The most abundant transcripts encoded fatty acid binding proteins, C-type lectins and activation associated secreted proteins, indicative of the diversity of functions that occur in this complex organ. Of particular interest to this study were the contigs that encoded for cysteine and metalloproteases, expanding the list of potential N. americanus haemoglobinases. In the N. americanus cDNA library, four contigs encoding for cathepsin B cysteine proteases were identified. Three contigs from the A. caninum and one contig from the N. americanus cDNA libraries encoded for metalloproteases, including astacin-like and O-sialoglycoprotein endopeptidases, neither of which had previously been reported from adult hookworms. Apart from haemoglobinases, other mRNAs encoding potential vaccine candidate molecules were identified, including anti-clotting factors, defensins and membrane proteins. This study confirmed that the gut of hookworms encodes a diverse range of proteases, some of which are likely to be involved in Hb digestion and have the potential to be hidden (cryptic) vaccine antigens. Four cysteine proteases (Na-CP-2, -3, -4 and -5) were identified from the gut cDNA library of N. americanus. All four proteases belong to the clan CA, family C1, share homology with human cathepsin B and possess a modified occluding loop. Real-time PCR indicated that all transcripts were up-regulated in the adult stage of the hookworm parasite with high levels of mRNA expression detected in gut cDNA. All four proteases were expressed in recombinant form, but only Na-CP-3 was successfully expressed in soluble form in the yeast Pichia pastoris. Proteolytic activity for Na-CP-3 was detected on a gelatin zymogen gel, however no catalytic activity was detected against the class-specific fluorogenic peptides Z-Phe-Arg-AMC and Z-Arg-Arg-AMC. Mass spectrometry analysis of the purified protein suggested that the pro-region had not been processed in trans when the protein was secreted by yeast. Incubation of Na-CP-3 in salt buffers containing dextran sulfate resulted in autoprocessing of the pro-region as detected by Western blot and catalytic activity was detected against Z-Phe-Arg-AMC. Activated Na-CP-3 did not digest intact tetrameric human Hb. The other three cysteine proteases (Na-CP-2, -4, and -5) were expressed in insoluble form in Escherichia coli. Antibodies to all four proteins (Na- CP-2 to 5) immunolocalised to the gut region of the adult worm, supporting mRNA amplification results and strongly indicated that they might play a role in nutrient acquisition. Hb digestion in blood feeding parasites such as schistosomes and Plasmodium spp. occurs via a semi-ordered cascade of proteolysis involving numerous enzymes. In Plasmodium falciparum, at least three distinct mechanistic classes of endopeptidases have been implicated in this process, and at least two classes have been implicated in schistosomes. A similar process is thought to occur in hookworms. An aspartic protease, Na-APR-1, was expressed in P. pastoris and purified protein was shown to cleave the class-specific fluorogenic peptide 7- Methoxycoumarin-4-Acetyl-GKPILFFRLK(DNP)-D-Arg-Amide. Recombinant Na- APR-1 was able to cleave intact human Hb and completely degrade the 16 kDa monomer and 32 kDa dimer within one hour. Recombinant Na-CP-3 was not able to cleave intact Hb, but was able to further digest globin fragments that had previously been digested with Na-APR-1. A clan MA metalloprotease, Na-MEP-1, was identified in gut tissue of N. americanus and was expressed in recombinant form in Hi5 insect cells using the baculovirus expression system. Recombinant Na-MEP-1 displayed proteolytic activity when assessed by gelatin zymography, but was incapable of cleaving intact Hb. However, Na-MEP-1 did cleave globin fragments which had previously been incubated with Na-APR-1 and Na-CP-3. Hb digested with all three proteases was subjected to reverse phase HPLC and peptides were analysed using Liquid Chromatography-Mass Spectrometry (LC-MS). A total of 74 cleavage sites were identified within Hb ƒ¿ and ƒÀ chains. Na-APR-1 was responsible for cleavage of Hb at the hinge region, probably unravelling the molecule so that Na- CP-3 and Na-MEP-1 could gain access to globin peptides. All three proteases were promiscuous in their subsite specificities, but the most common P1-P1Œ residues were hydrophobic and/or bulky in nature, such as Phe, Leu and Ala. Antibodies to all three proteins (Na-APR-1, -CP-3, -MEP-1) immunolocalised to the gut region of the worm, further supporting their roles in Hb degradation. These results suggest that Hb degradation in N. americanus follows a similar pattern to that which has been described in Plasomdium falciparum. Studies conducted in this project have identified a number of potential haemoglobinases and have demonstrated that the gut region of the hookworm contains a multitude of proteases which could be targeted for production of new chemotherapies or as vaccine candidates. Results presented here also suggest that the Hb degradation process occurs in an ordered cascade, similar to those which have been reported in other haematophagous parasites. More importantly, it has been confirmed that Na-APR-1 plays a crucial role in the initiation of the Hb degradation process and therefore targeting this molecule as a vaccine candidate could provide high levels of protection against hookworm infection.
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Gonzalez, Santana Bibiana. "Cysteine proteases: potential serodiagnostic reagents for human Schistosomiasis and Fasciolosis." Thesis, McGill University, 2012. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=110691.
Повний текст джерелаАнотація:
Schistosomiasis and fasciolosis are parasitic diseases that affect a great number of people, particularly in developing countries, and cause huge global morbidity. Diagnosis is essential for control, treatment and prognosis of the diseases and yet a simple, cheap, sensitive and specific assay is not readily available for either of them. In the current study, cathepsin B (SmCB) and cathepsin L1 (FhCL1) were investigated as potential diagnostic reagents to detect schistosomiasis and fasciolosis, respectively, in humans. The genes encoding SmCB and FhCL1 were expressed Pichia pastoris and the proteins isolated to homogeneity by affinity chromatography. The SmCB ELISA was optimized for antigen concentration, primary antibody dilution and secondary antibody dilution using a pool of sera obtained from patients that were coprologically-positive or negative for schistosomiasis. A clear distinctive was achieved between these two sera pools. However, when employed to screen a panel of patients from Senegal the test failed to provide satisfactory discrimination between schistosoma-infected and schistosoma-negative individuals. The FhCL1 ELISA was optimized using sera from Fasciola-infected individuals from Cuba, and samples from Cuban and Canadian non-infected patients. We determined the optimal dilution for the primary antibody and also assessed/compared the performance of anti-total IgG, IgG4, IgG1 and IgG2 secondary conjugated antibodies. Total IgG provided the best discrimination between Fasciola- infected and non-Fasciola infected individuals with a 99.99% sensitivity and specificity. Furthermore, by screening sera obtained from patients infected with various worm and protozoan diseases we showed that the FhCL1 ELISA does not cross-react with other diseases commonly found in similar geographical regions as fasciolosis. In conclusion, diagnosis of human schistosomiasis still remains uncertain and more studies need to be performed to improve our diagnostic test using SmCB. On the other hand, we have developed a simple, sensitive, specific and accurate test to detected human fasciolosis by using FhCL1, a major protease released by the parasite. The P. pastoris expression system allowed us to obtain up to 80 mg of FhCL1 enzyme per 4 L culture. Therefore, we not only have developed a standardized test that showed high specificity and sensitivity but we also have the methodology to obtain sufficent quantities of antigen needed future mass screening of human fasciolosis in affected regions.La schistosomiase et la fasciolose sont deux maladies parasitaires qui touchent un grand nombre de personnes, en particulier dans les pays en développement, causant une morbidité élevée. Le diagnostic est essentiel pour le contrôle, le traitement et le pronostic de ces maladies et pourtant aucun essai simple, abordable, sensible et spécifique n'est disponible à ce jour pour l'une d'entre elles. Dans le cadre de la présente étude, cathepsine B (SmCB) et cathepsine L1 (FhCL1) ont fait l'objet d'une investigation sur leur potentiel à être utiliser pour diagnostiquer la schistosomiase et fasciolose, respectivement, chez les humains. Dans la présente étude, les gènes encodant SmCB et FhCL1 ont été exprimés dans Pichia pastoris et les protéines isolées par chromatographie d'affinité. Le test ELISA pour SmCB a été optimisé pour une concentration en antigène et pour une dilution d'anticorps primaire et secondaire en utilisant un pool de sérums provenant de patients qui étaient positifs ou négatifs pour la schistosomiase suivant des examens coprologique. Une distinction claire entre ces deux bassins de sérums a été observée. Toutefois, lorsque le test a été utilisé pour dépister des patients du Sénégal, il a échoué à fournir une discrimination satisfaisante entre les individus infectés et non-infectés par la schistosomiase.Le test ELISA pour FhCL1 a été optimisé à l'aide de sérums provenant de personnes cubaines infectées par la fasciolose et de patients non-infectés de Cuba et du Canada. Nous avons déterminé la dilution optimale pour l'anticorps primaire et également évalué et comparé la performance des anticorps secondaires conjugués contre les IgG totaux, IgG4, IgG1 et IgG2. Les IgG totaux ont fourni la meilleure discrimination entre les personnes infectées et non-infectées par la fasciolose avec une sensibilité et une spécificité de 99.99 %. En plus, en appliquant le test de dépistage sur des patients infectés par d'autres variétés de vers et de protozoaires, nous avons démontré que le test ELISA pour FhCL1 ne réagit pas de façon croisée avec d'autres maladies retrouvées couramment dans les régions géographiques où se trouve la fasciolose.En conclusion, le diagnostic de la schistosomiase humaine reste encore incertain et d'autres études doivent être effectuées pour améliorer notre test de dépistage utilisant SmCB. D'autre part, nous avons développé un test simple, sensible, spécifique et précis pour le dépistage de la fasciolose humaine en utilisant FhCL1, une protéase majeure relâchée par le parasite. Le système d'expression de P. pastoris nous a permis d'obtenir jusqu'à 80 mg de FhCL1 par 4 litres de culture. Dans la présente étude, nous avons non seulement mis au point un test standardisé démontrant une spécificité et une sensibilité élevées, mais également développé une procédure pour produire de grandes quantités d'antigènes nécessaires au dépistage à grande échelle de la fasciolose humaine dans les régions touchées.
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Riddick, Antony C. P. "Expression profiling of proteases and related genes in human prostate tumours." Thesis, University of East Anglia, 2002. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.398820.
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Gast, Alain. "Proteases et emphyseme pulmonaire : etude des inhibiteurs de proteases recueillis par lavage bronchoalveolaire." Université Louis Pasteur (Strasbourg) (1971-2008), 1987. http://www.theses.fr/1987STR13070.
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Wang, Bingjie. "Novel function of human beta-defensin 2 : protecting epidermal barrier against pathogenic proteases." Thesis, University of Edinburgh, 2017. http://hdl.handle.net/1842/28756.
Повний текст джерелаАнотація:
Atopic Dermatitis (AD) is a common chronic relapsing inflammatory skin disease affecting 15 - 20% of children and 2 - 10% of adults worldwide, with significant morbidity. A hallmark of AD is disruption of the critical barrier function of upper epidermal layers, causatively linked to environmental stimuli, genetics and infections. Another typical feature of AD is skin infections, especially from Staphylococcus aureus (S. aureus), which closely relates with the disease severity. Although not a normal flora, S. aureus is found on 75-100% of AD lesions (< 30% on healthy skin). S. aureus secrete a range of virulence factors, including extracellular toxins and proteases which contribute to disease pathogenesis. S. aureus serine protease A (SspA/V8) is a well-characterised extracellular protease widely expressed among different S. aureus strains. The pathogenic effect of V8 protease has been demonstrated in vivo, damaging murine skin integrity via effects on the stratum corneum (SC), but the targets for this V8-mediated damage remains unclear. The capacity of proteases to induce barrier dysfunction has been proposed as a key driving force in the initiation and exacerbation of AD. Thus, understanding the host factors that maintain barrier function is a priority in developing novel therapeutic approaches. This thesis therefore aimed at detecting host factors which can combat the barrier dysfunction caused by pathogenic proteases, assessing their relevance in vitro and ex vivo and elucidating the underlying mechanisms. Firstly, an in vitro skin barrier integrity model was developed, using both immortalized and primary keratinocytes, to evaluate the barrier damage mediated by pathogenic proteases. The results revealed that S. aureus protease SspA/V8 is the dominant secreted factor (in laboratory and AD clinical strains of S. aureus) inducing barrier integrity impairment. In addition, studies demonstrated that V8 protease itself was sufficient to induce barrier disruption, and this phenotype was not dependent on cell death, but rather on breaking down of cell-cell junctions. Key tight junction proteins including claudin-1 and occludin were found to be degraded by V8 protease. Next, a wide range of host and bacterial factors were investigated to determine whether they could promote protection of keratinocytes against V8 damage. Several factors, including IL-1β, TNF-α, heat-killed Staphylococcus epidermidis (which is the main skin normal flora), were found to induce protection against V8 protease, with IL-1β having the strongest effect. In addition, data indicated that this IL-1β-mediated protection was independent of effects on claudin-1 but occurred via secretion of a transferrable host factor. Induction of keratinocyte expression of the antimicrobial/host defence peptide human beta-defensin 2 (hBD2) was found to be the mechanism underpinning this IL-1β- induced protective effect. Endogenous hBD2 expression was required and sufficient for protection against V8 protease-mediated integrity damage, and exogenous application of hBD2 was also protective. An ex vivo model using human skin tissue was also established to address this novel function of hBD2, and preliminary data indicated that exogenous hBD2 protected against V8-mediated damage in this system. Overall, my data reveal a novel function for the antimicrobial/host defence peptide hBD2. This modulatory property of hBD2, independent of its antibacterial effects, gives new significance to the defective induction of hBD2 in the barrier-defective skin lesions of AD and indicates therapeutic potential to prevent S. aureus-mediated aggravation of skin barrier dysfunction in AD.
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West, Andrew. "Investigations by mass spectrometry of the interactions of novel serine protease inhibitors with herpes simplex virus type 2 and human cytomegalovirus proteases." Thesis, University of Warwick, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.343830.
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Mulloy, Rory. "Identification of Transmembrane and Extracellular Host Proteases that Promote Human CoV Entry and Syncytium Formation." Thesis, Université d'Ottawa / University of Ottawa, 2021. http://hdl.handle.net/10393/42673.
Повний текст джерелаКниги з теми "Human proteases"
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Chakraborti, Sajal, Tapati Chakraborti, and Naranjan S. Dhalla, eds. Proteases in Human Diseases. Singapore: Springer Singapore, 2017. http://dx.doi.org/10.1007/978-981-10-3162-5.
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Cheronis, John Chris Dion, 1951- and Repine John E, eds. Proteases, protease inhibitors, and protease-derived peptides: Importance in human pathophysiology and therapeutics. Basel: Birkhäuser Verlag, 1993.
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West, A. Investigations by mass spectrometry of the interactions of novel serine protease inhibitors with Herpes Simplex Virus type 2 and Human Cytomegalovirus proteases. [s.l.]: typescript, 1999.
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Genesio, Murano, ed. Protease inhibitors of human plasma: Biochemistry and pathophysiology. Westbury, N.Y: PJD Publications, 1986.
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Massimi, Isabella. SspB cysteine protease of Staphylococcus aureus promotes detachment of human keratinocytes and degrades fibronectin and vitronectin. Ottawa: National Library of Canada, 2001.
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John Chris Dion Cheronis (Editor) and John E. Repine (Editor), eds. Proteases, Protease Inhibitors & Proteasederived Peptides : Importance in Human... Birkhauser Boston, 1993.
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Dhalla, Naranjan S., Sajal Chakraborti, and Tapati Chakraborti. Proteases in Human Diseases. Springer, 2018.
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Dhalla, Naranjan S., Sajal Chakraborti, and Tapati Chakraborti. Proteases in Human Diseases. Springer, 2017.
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Dhalla, Naranjan S., Sajal Chakraborti, and Tapati Chakraborti. Proteases in Human Diseases. Springer, 2017.
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Dhalla, Naranjan S., Sajal Chakraborti, and Tapati Chakraborti. Proteases in Human Diseases. Springer, 2017.
Знайти повний текст джерелаЧастини книг з теми "Human proteases"
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Whalley, E. T., and J. C. Cheronis. "Kinin Antagonists as Human Therapeutics." In Proteases, Protease Inhibitors and Protease-Derived Peptides, 167–76. Basel: Birkhäuser Basel, 1993. http://dx.doi.org/10.1007/978-3-0348-7397-0_14.
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Patton, Lavonne M. "In Vivo Evaluation of MDL 201,404YA, A Novel Inhibitor of Human Neutrophil Elastase." In Proteases, Protease Inhibitors and Protease-Derived Peptides, 83–96. Basel: Birkhäuser Basel, 1993. http://dx.doi.org/10.1007/978-3-0348-7397-0_7.
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Kirschenheuter, Gary P., Josef Oleksyszyn, Lyle W. Spruce, Maciej Wieczorek, Thomas M. Kloppel, Sanford R. Simon, and John C. Cheronis. "Synthesis and Characterization of Human Neutrophil Elastase Inhibitors Derived from Aromatic Esters of Phenylalkanoic Acids." In Proteases, Protease Inhibitors and Protease-Derived Peptides, 71–82. Basel: Birkhäuser Basel, 1993. http://dx.doi.org/10.1007/978-3-0348-7397-0_6.
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Sarkar, Abhijit, Sumit Ghosh, Sayanta Dutta, and Parames C. Sil. "Proteases in Neuropathophysiology." In Proteases in Human Diseases, 131–45. Singapore: Springer Singapore, 2017. http://dx.doi.org/10.1007/978-981-10-3162-5_7.
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Ranogajec, Irena. "Matrix Metalloproteinases in Breast Carcinoma." In Proteases in Human Diseases, 3–20. Singapore: Springer Singapore, 2017. http://dx.doi.org/10.1007/978-981-10-3162-5_1.
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Rachel, K. Vijaya, and Gandreddi V. D. Sirisha. "Serine Proteases and Their Inhibitors in Human Health and Disease." In Proteases in Human Diseases, 195–226. Singapore: Springer Singapore, 2017. http://dx.doi.org/10.1007/978-981-10-3162-5_10.
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Chakraborty, Sibani, Asim K. Bera, Ankur Chaudhuri, and Satyajit Sen. "Metalloproteases and Human Diseases: The Astacin Family." In Proteases in Human Diseases, 227–45. Singapore: Springer Singapore, 2017. http://dx.doi.org/10.1007/978-981-10-3162-5_11.
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Tandon, Veena, Bidyadhar Das, and Shakti Kumar. "Proteases of Parasitic Helminths: Their Metabolic Role in Establishment of Infection in the Host." In Proteases in Human Diseases, 247–62. Singapore: Springer Singapore, 2017. http://dx.doi.org/10.1007/978-981-10-3162-5_12.
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Rustagi, Yashika, Aditi Jain, Sharad Saxena, and Vibha Rani. "Natural Polyphenols as Prospective Inhibitors for MMPs Remodeling in Human Diseases." In Proteases in Human Diseases, 263–83. Singapore: Springer Singapore, 2017. http://dx.doi.org/10.1007/978-981-10-3162-5_13.
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Pramanik, Pijush Kanti, Dibyendu Paik, Asmita Pramanik, Md Nur Alam, Partha Das, and Tapati Chakraborti. "Autophagic Proteases: Functional and Pathophysiological Aspects." In Proteases in Human Diseases, 285–301. Singapore: Springer Singapore, 2017. http://dx.doi.org/10.1007/978-981-10-3162-5_14.
Повний текст джерелаТези доповідей конференцій з теми "Human proteases"
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Farache Trajano, Luiza, Rebecca Moore, and Quentin Sattentau. "The Presence of Chemical Cross-Linking Stabilises HIV-1 Envelope Glycoprotein Trimer Antigens in a Model of Intramuscular Immunisation." In Building Bridges in Medical Science 2021. Cambridge Medicine Journal, 2021. http://dx.doi.org/10.7244/cmj.2021.03.001.4.
Повний текст джерелаАнотація:
Background: The HIV-1 envelope glycoprotein (Env) is the target of antigen design for antibody- based vaccination. In 2019, four trimeric Env vaccines entered an experimental trial: ConM, ConS, and their cross-linked counterparts. The trimers were formulated with MPLA adjuvant. Studies have demonstrated that adjuvants trigger neutrophil infiltration. Neutrophils activate and degranulate releasing proteases, namely elastase and cathepsinG. Aims: To assess the stability and immunogenicity of these vaccines in the presence of adjuvant- recruited neutrophils and their proteolytic enzymes. Methods: Trimers were incubated with commercially-sourced proteases. To analyse stability, samples were reduced, denatured and separated using gel electrophoresis. To assess antibody binding, a trimer-protease incubation was followed by an ELISA. To establish more physiologically relevant conditions, harvested neutrophils were exposed to various adjuvants. The supernatant, shown to contain elastase, was incubated alongside the vaccines. The reducing and denaturing gels, as well as the ELISA, was repeated. Results: Gel analysis revealed that un-crosslinked trimers underwent significant digestion whereas cross-linking conferred enhanced stability. In the presence of neutrophil-sourced protease-containing-supernatant, trimers displayed resistance to digestion. The differential stability profile of Env trimers when exposed to commercially sourced compared to supernatant- derived proteases may be due to the inhibitory effect of human serum on elastase. Antibody epitopes were maintained in vitro. Conclusion: The vaccine antigens are sensitive to enzymatic degradation. This is reduced by cross-linking and human serum.
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Kindel, G., and J. Fareed. "MODULATORY EFFECT OF SERINE PROTEASES AND RELATED ENZYMES ON ISOLATED SMOOTH MUSCLE PREPARATIONS." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644602.
Повний текст джерелаАнотація:
Thrombin and related proteases produce varying pharmacologic responses in animal models. To more specifically study the in vivo actions of thrombin and related proteases, we have used isolated tissue preparations of the rabbit aortic strip (RAS), isolated guinea pig ileum (GPI) and isolated rat uterus (RU). Standard tissue-agonist regimens include epinephrine, thromboxane B2 with RAS; bradykinin, acetylcholine, histamine and serotonin with GPI; and acetylcholine, bradykinin and angiotensin with RU. The smooth muscle modulant action of numerous proteinases were screened in these regimens by bracketing the median dose response of the individual agonists. Protease complexes such as serum (rabbit, human and guinea pig), activated and nonactivated prothrombin complex concentrates and pancreatin were shown to produce varying but similar contractile responses as obtained by the standard agonists. Sera produced a dose-dependent contraction of the RAS, GPI and RU preparations. Various forms of thrombin produced different degrees of contraction of RAS accompanied by a desensitization process. On a molar basis the order of contractile activity ranged α > β>γ > nitro > DIP. All thrombins were found to augment the epinephrine and thromboxane B2 induced contraction of the RAS. Bovine and human factor Xa produced marked dilatation of the RAS but did not have any effect on the GPI and RU preparations. These results suggest that proteases exert direct musculotropic actions on smooth muscles. This should be taken into consideration in the pathophysiology of vascular spasms.
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Tavakolinejad, Alireza, and Vinod Suresh. "Activity of ENaC-activating serine proteases in human alveolar epithelial cells." In ERS International Congress 2020 abstracts. European Respiratory Society, 2020. http://dx.doi.org/10.1183/13993003.congress-2020.1901.
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Wasi, S., S. Juodvalkis, P. Alles, and J. E. Aubin. "STUDIES ON THE DIRECT PROTEOLYTIC ACTION OF HUMAN TISSUE PLASMINOGEN ACTIVATOR ON HUMAN FIBRONECTIN AND VITRONECTIN." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644376.
Повний текст джерелаАнотація:
The ability of cells to make or break specific attachments to extracellular matrix (ECM) and other cells is important in cell migration, proliferation and wound repair. Specific attachment proteins believed to be involved in mediating these interactions comprise functional domains joined by protease sensitive segments. Proteases can conceivably modulate cellular interactions by releasing functional domains from parent molecules. Tissue plasminogen activator (t-pA) is known to participate in various pathophysiological processes. That t-pA may also act directly on structural proteins has not been investigated. We have studied the direct proteolytic action of melanoma t-pA on fibronectin (FN), vitronectin (VN) and laminin (LN). These were incubated with t-pA for 0 to 48 h in 50 mM Tris HCi, pH 7.4. The cleavage products were separated on polyacrylamide slab gels and blotted onto nitrocellulose strips. FN and VN fragments with cell attachment properties were identified by incubating the strips with human gingiva fibroblasts and staining with Amido black. Monoclonal antibodies to FN were used to identify heparin, cell and gelatin binding fragments. VN was converted to a major 55 Kd product as a function of time. Lower molecular weight species migrating at 45 Kd, 30 Kd and 15 Kd positions were also identified. Most of these fragments possessed cell attachment properties. LN became susceptible to t-pA digestion after dénaturation with H2O2. The catalytic activity of t-pA could be inhibited in the presence of nitrophenyl-p-guinidino benzoate (a synthetic inhibitor of plasminogen activator), whereas O-phenanthroline (a metalloprotease inhibitor), α 2-antiplasmin and trasylol had no effect. A monoclonal IgG preparation (HI 72 A1, kindly provided by Dr. David J. Loskutoff) that specifically inhibits t-pA also inhibited the protelyotic action of t-pA on FN. These data suggest that direct proteolytic action of t-pA on adhesive proteins may modulate cellular behaviour in various normal and pathological conditions which involve dynamic interactions between cells and ECM and where plasminogen activator levels are elevated either transiently or permanently, for example during tissue remodelling, wound-related repair and thrombolytic therapy.
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Tucker, Torry A., Ann Jeffers, Alexia Alvarez, Kathleen Koenig, L. Vijaya M. Rao, and Steven Idell. "Coagulation Cascade Proteases Induce Mesenchymal Transition In Human Pleural Mesothelial Cells: Implications For Pleural Fibrosis." In American Thoracic Society 2012 International Conference, May 18-23, 2012 • San Francisco, California. American Thoracic Society, 2012. http://dx.doi.org/10.1164/ajrccm-conference.2012.185.1_meetingabstracts.a5568.
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Yamaguchi, Kosuke, Hiroki Hikumi, Shinji Matsumaga, Miyako Takata, Naoki Kinoshita, Kiyoshi Hashimoto, Masaki Nakamoto, et al. "Abstract 454: ADAM proteases shed UL-16-binding protein 2 in human lung cancer cell lines." In Proceedings: AACR 104th Annual Meeting 2013; Apr 6-10, 2013; Washington, DC. American Association for Cancer Research, 2013. http://dx.doi.org/10.1158/1538-7445.am2013-454.
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Han, Guangchun, Ansam Sinjab, Kieko Hara, Warapen Treekitkarnmongkol, Patrick Brennan, Kyle Chang, Elena Bokatenkova, et al. "Abstract 702: Single-cell expression landscape of SARS-CoV-2 receptorACE2and host proteases in human lung adenocarcinoma." In Proceedings: AACR Annual Meeting 2021; April 10-15, 2021 and May 17-21, 2021; Philadelphia, PA. American Association for Cancer Research, 2021. http://dx.doi.org/10.1158/1538-7445.am2021-702.
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Kasaian, MT, A. Margulis, KH Nocka, B. Deng, M. Fleming, and SJ Goldman. "Mast Cell-Dependent Contraction of Human Airway Smooth Muscle Cells: Influence of Cytokines, Matrix Metalloproteases, and Serine Proteases." In American Thoracic Society 2009 International Conference, May 15-20, 2009 • San Diego, California. American Thoracic Society, 2009. http://dx.doi.org/10.1164/ajrccm-conference.2009.179.1_meetingabstracts.a3706.
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Oldenburg, Emil, Christine R. Schar, Eva Lange, Terry F. Plasse, Danielle T. Abramson, Reza Fathi, Eric M. Towler, Mark Levitt, and Jan K. Jensen. "Abstract 4200: New potential therapeutic applications of WX-UK1, as a specific and potent inhibitor of human trypsin-like proteases." In Proceedings: AACR Annual Meeting 2018; April 14-18, 2018; Chicago, IL. American Association for Cancer Research, 2018. http://dx.doi.org/10.1158/1538-7445.am2018-4200.
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Furmaniak-Kazmierczak, E., J. Jagielski, and T. Wilusz. "THE EFECT OF CMTI-I INHIBITOR ON HUMAN BLOOD CLOTTING SYSTEM." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644327.
Повний текст джерелаАнотація:
Polipeptyde inhibitors for different serine proteases have been isolated from a variety of plants. Among them there are the inhibitors from squash seeds of molecular weight about 3 300 /1/. The experiments were carried out to determine the effect of one of the squash inhibitors /CMTI-I/ on human blood clotting system. The 0.1 ml of inhibitor /O,8-100 ug/ was added to 0,1 ml of normal intact plasma and incubated 0,5, 15, 30 and 60 minutes at 37°C. It was found that partial tromboplastin time /PTT/ and activated partial thromboplastin time /APTT/ were prolonged. CMTI-I did not show a progressive mode of action upon prolonged time of incubation. There was no effect of CMTI-I on prothrombin time /PT/, thrombin time /TT/ and Stypven-cephalin time /ScT/. The influence of CMTI-I on APTT of factor-XII and factor-XI deficient plasmas as well as on a plasma without factor-XII and factor-XI /exhausted plasma/ was studied. The APTTs of the factor-XII and factor-XI deficient plasmas were prolonged while the APTT of the "exhausted plasma" was unchanged. The performed experiments shown that CMTI-I inhibitor blocks the clot-promoting activity of contact activated plasma. This inhibitory action is stronger in the case of factor XI than of factor XII.1. Wieczorek M., et al., 1985, Biochem. Biophys. Res. Commun. 126:646-652.
Звіти організацій з теми "Human proteases"
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Gross, Clark L., Juanita J. Guzman, Charlene M. Corun, Marian R. Nelson, and William J. Smith. Measurement of Protease Release by a Fluorogenic Casein Assay in Human Cells Exposed In Vitro to Sulfur Mustard. Fort Belvoir, VA: Defense Technical Information Center, October 2000. http://dx.doi.org/10.21236/ada390636.
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Ohad, Itzhak, and Himadri Pakrasi. Role of Cytochrome B559 in Photoinhibition. United States Department of Agriculture, December 1995. http://dx.doi.org/10.32747/1995.7613031.bard.
Повний текст джерелаАнотація:
The aim of this research project was to obtain information on the role of the cytochrome b559 in the function of Photosystem-II (PSII) with special emphasis on the light induced photo inactivation of PSII and turnover of the photochemical reaction center II protein subunit RCII-D1. The major goals of this project were: 1) Isolation and sequencing of the Chlamydomonas chloroplast psbE and psbF genes encoding the cytochrome b559 a and b subunits respectively; 2) Generation of site directed mutants and testing the effect of such mutation on the function of PSII under various light conditions; 3) To obtain further information on the mechanism of the light induced degradation and replacement of the PSII core proteins. This information shall serve as a basis for the understanding of the role of the cytochrome b559 in the process of photoinhibition and recovery of photosynthetic activity as well as during low light induced turnover of the D1 protein. Unlike in other organisms in which the psbE and psbF genes encoding the a and b subunits of cytochrome b559, are part of an operon which also includes the psbL and psbJ genes, in Chlamydomonas these genes are transcribed from different regions of the chloroplast chromosome. The charge distribution of the derived amino-acid sequences of psbE and psbF gene products differs from that of the corresponding genes in other organisms as far as the rule of "positive charge in" is concerned relative to the process of the polypeptide insertion in the thylakoid membrane. However, the sum of the charges of both subunits corresponds to the above rule possibly indicating co-insertion of both subunits in the process of cytochrome b559 assembly. A plasmid designed for the introduction of site-specific mutations into the psbF gene of C. reinhardtii. was constructed. The vector consists of a DNA fragment from the chromosome of C. reinhardtii which spans the region of the psbF gene, upstream of which the spectinomycin-resistance-conferring aadA cassette was inserted. This vector was successfully used to transform wild type C. reinhardtii cells. The spectinomycin resistant strain thus obtained can grow autotrophically and does not show significant changes as compared to the wild-type strain in PSII activity. The following mutations have been introduced in the psbF gene: H23M; H23Y; W19L and W19. The replacement of H23 involved in the heme binding to M and Y was meant to permit heme binding but eventually alter some or all of the electron transport properties of the mutated cytochrome. Tryptophane W19, a strictly conserved residue, is proximal to the heme and may interact with the tetrapyrole ring. Therefore its replacement may effect the heme properties. A change to tyrosine may have a lesser affect on the potential or electron transfer rate while a replacement of W19 by leucine is meant to introduce a more prominent disturbance in these parameters. Two of the mutants, FW19L and FH23M have segregated already and are homoplasmic. The rest are still grown under selection conditions until complete segregation will be obtained. All mutants contain assembled and functional PSII exhibiting an increased sensitivity of PSII to the light. Work is still in progress for the detailed characterization of the mutants PSII properties. A tobacco mutant, S6, obtained by Maliga and coworkers harboring the F26S mutation in the b subunit was made available to us and was characterized. Measurements of PSII charge separation and recombination, polypeptide content and electron flow indicates that this mutation indeed results in light sensitivity. Presently further work is in progress in the detailed characterization of the properties of all the above mutants. Information was obtained demonstrating that photoinactivation of PSII in vivo initiates a series of progressive changes in the properties of RCII which result in an irreversible modification of the RCII-D1 protein leading to its degradation and replacement. The cleavage process of the modified RCII-D1 protein is regulated by the occupancy of the QB site of RCII by plastoquinone. Newly synthesized D1 protein is not accumulated in a stable form unless integrated in reassembled RCII. Thus the degradation of the irreversibly modified RCII-D1 protein is essential for the recovery process. The light induced degradation of the RCII-D1 protein is rapid in mutants lacking the pD1 processing protease such as in the LF-1 mutant of the unicellular alga Scenedesmus obliquus. In this case the Mn binding site of PSII is abolished, the water oxidation process is inhibited and harmful cation radicals are formed following light induced electron flow in PSII. In such mutants photo-inactivation of PSII is rapid, it is not protected by ligands binding at the QB site and the degradation of the inactivated RCII-D1 occurs rapidly also in the dark. Furthermore the degraded D1 protein can be replaced in the dark in absence of light driven redox controlled reactions. The replacement of the RCII-D1 protein involves the de novo synthesis of the precursor protein, pD1, and its processing at the C-terminus end by an unknown processing protease. In the frame of this work, a gene previously isolated and sequenced by Dr. Pakrasi's group has been identified as encoding the RCII-pD1 C-terminus processing protease in the cyanobacterium Synechocystis sp. PCC 6803. The deduced sequence of the ctpA protein shows significant similarity to the bovine, human and insect interphotoreceptor retinoid-binding proteins. Results obtained using C. reinhardtii cells exposes to low light or series of single turnover light flashes have been also obtained indicating that the process of RCII-D1 protein turnover under non-photoinactivating conditions (low light) may be related to charge recombination in RCII due to back electron flow from the semiquinone QB- to the oxidised S2,3 states of the Mn cluster involved in the water oxidation process.