Dissertations / Theses: 'Secretory (E/S) proteins'

1

Shukeir, Nicholas. "Molecular mechanism(s) of prostate cancer progression : potential of therapeutic modalities." Thesis, McGill University, 2009. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=115853.

Full text

Abstract:

Prostate cancer remains one of the most commonly diagnosed cancers in men and is a leading cause of cancer death. While great success has been achieved at curing early stage prostate cancer, limited success has been obtained when treating late-stage hormone independent prostate cancer. This is due to the increased propensity for skeletal and non-skeletal metastases. Thus development of novel effective therapeutic modalities against late stage prostate cancer is of critical importance.
Towards these objectives, I have focused my attention on the role of prostate secretory protein (PSP-94) which is expressed in normal individuals and in patients with early stage prostate cancer. Using our well established in vivo models of prostate cancer, I have evaluated the ability of PSP-94 and its amino acids 31-45 required (PCK3145) to decrease tumor growth and skeletal metastases in vivo and evaluated the potential mechanism(s) associated with PCK3145 anti-cancer actions.
Prostatic cancer can also develop as a result of epigenetic activation of tumor promoting genes. To evaluate the role of methylation in prostate cancer, late stage prostate cancer cells were treated with the universal methylating agent S-adenosylmethionine (SAM) and an anti-sense oligonucleotide directed against MBD2 (AS). Scrambled oligonucleotide was included as a control (S). Both SAM and MBD2-AS resulted in inhibition in uPA, MMP-2 and VEGF production leading to decreased tumor cell invasive capacity. However, SAM and MBD2-AS were not able to either further repress partially methylated genes (GSTP1) or reactivate already methylated genes (AR). Furthermore, SAM and MBD2-AS treatment resulted in significant reduction in tumor growth in vivo . Immunohistochemical and RT-PCR analyses carried out on SAM and MBD2-AS tumors revealed decreased protein and mRNA expression of uPA and MMP-2 which was partially due to increased methylation of the respective promoters even after 10 weeks post in vitro treatment as analyzed by bisulfate sequencing. In addition decreased levels of angiogenesis and tumor survival markers were observed.
Collectively, these studies are aimed at the development of novel reliable approached to diagnose and treat advanced, hormone refractory prostate cancer to reduce tumor associated morbidity and mortality.

APA, Harvard, Vancouver, ISO, and other styles

2

Čiplys, Evaldas. "Analysis of maturation of measles virus hemaglutinin in yeast S. cerevisiae and P. pastoris secretory pathway and humanization of yeast cells." Doctoral thesis, Lithuanian Academic Libraries Network (LABT), 2011. http://vddb.laba.lt/obj/LT-eLABa-0001:E.02~2011~D_20111227_092012-09066.

Full text

Abstract:

The aims of the study were to determine the reasons for unsuccessful expression of measles virus hemaglutinin (MeH) in the yeast cells and to generate a stable yeast strains with integrated genes of protein secretory pathway of human cells and to examine influence of coded human proteins on MeH maturation. For the firs time, overexpression of MeH in yeast S. cerevisiae and P. pastoris was described. It was demonstrated that mechanisms of cotranslational translocation into the endoplasmic reticulum (ER) and protein maturation in the ER of yeast cells are not adapted to deal with for such complex virus glycoproteins. Proteomic analysis revealed, that overexpression of human virus surface protein precursors induces cytosolic unfolded protein response (UPR-cyto) in the yeast S. cerevisiae. A key feature of this response is the formation of extremely large aggregates involving macromolecular structures of eEF1A. Efficient mammalian like cotranslational translocation pathway was attempted to reconstitute in yeast cells by transferring human SRP, Sec61 complexes and TRAM1 protein. Human chaperones BiP, clanexin, calreticulin, ERp57 and PDI were transferred to the yeast cells to create suitable environment for maturation of MeH in the ER. Even though yeast strains, able to produce biologically active MeH protein, were not generated during this study, results show, that humanization of yeast secretory pathway, designed for producing active virus glycoproteins, is possible.
Baigiamojo darbo tikslai – nustatyti neefektyvios žmogaus virusų glikobaltymų raiškos mielėse priežastis ir sukurti mielių kamienus su integruotais žmogaus ląstelių sekrecinio kelio genais bei ištirti jų įtaką glikobaltymų sintezei ir brendimui mielėse. Darbo eigoje pirmą kartą buvo aprašytos tymų viruso hemagliutinino (TVH) sintezės galimybės mielėse Saccharomyces cerevisiae ir Pichia pastoris. Parodyta, kad mielių ko-transliacinio baltymų perkėlimo į endoplazminį tinklą (ET) ir ET baltymų sulankstymo mechanizmai nėra pritaikyti sudėtingų virusinių baltymų brendimui, todėl klasikinės mielių rūšys ir standartiniai rekombinantinių baltymų raiškos ir gryninimo protokolai nėra tinkami diagnostikai ir vakcinų kūrimui reikalingo TVH baltymo gavimui. Proteominė S. cerevisiae ląstelių, sintetinančių TVH baltymą, analizė leido nustatyti kad, TVH sintezė mielėse sukelia neseniai literatūroje aprašytą citoplazminį nesusivyniojusių baltymų atsaką (UPR-cyto). Pagrindinis šiame darbe aprašyto atsako į stresą požymis yra ypatingai didelių baltymų agregatų, kurių šerdį sudaro TVH ir mielių eEF1A baltymai, susidarymas. Žmogaus tipo ko-transliacinį baltymų pernešimą į ET mielių ląstelėse bandyta atkurti perkeliant žmogaus SRP, Sec61 kompleksų ir TRAM1 baltymus, o siekiant sukurti tinkamas TVH baltymo brendimui sąlygas, mielių ląstelių ET buvo sintetinami pagrindiniai žmogaus ląstelių ET šaperonai – BiP, kalretikulinas, kalneksinas, PDI ir ERp57. Nors šiame darbe nepavyko sukurti mielių... [toliau žr. visą tekstą]

APA, Harvard, Vancouver, ISO, and other styles

3

Matiach, Alexei. "Retrograde protein trafficking of Emp47p in the early secretory pathway of S. cerevisiae a novel mutant screen uncovers the influence of oxidative stress on protein trafficking /." Kassel : Kassel Univ. Press, 2002. http://deposit.d-nb.de/cgi-bin/dokserv?idn=970323417.

Full text

APA, Harvard, Vancouver, ISO, and other styles

4

Matiach, Alexei [Verfasser]. "Retrograde protein trafficking of Emp47p in the early secretory pathway of S. cerevisiae : a novel mutant screen uncovers the influence of oxidative stress on protein trafficking / vorgelegt von Alexei Matiach." Kassel : Kassel Univ. Press, 2002. http://d-nb.info/970323417/34.

Full text

APA, Harvard, Vancouver, ISO, and other styles

5

Clark, Richard. "Sorting proteins to the secretory lysosome." Thesis, University of Oxford, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.404112.

Full text

APA, Harvard, Vancouver, ISO, and other styles

6

Drake, Lesley Jayne. "Characterization of execretory/secretory proteins of Trichuris." Thesis, Imperial College London, 1993. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.294921.

Full text

APA, Harvard, Vancouver, ISO, and other styles

7

Gomi, Masahiro, Ryusuke Sawada, Masashi Sonoyama, Shigeki Mitaku, 雅裕五味, 隆介澤田, 正史園山, and 成樹美宅. "Comparative proteomics of the prokaryota using secretory proteins." Chem-Bio Informatics Society, 2005. http://hdl.handle.net/2237/9270.

Full text

APA, Harvard, Vancouver, ISO, and other styles

8

Kemter, Andrea Maria. "Immunomodulatory proteins in Heligmosomoides polygyrus excretory/secretory products." Thesis, University of Edinburgh, 2016. http://hdl.handle.net/1842/23656.

Full text

Abstract:

Infections with parasitic helminths are counted as neglected tropical diseases; they infect millions of people worldwide, causing high morbidity and economic loss. Many parasites establish long lasting infections in the host by blocking immune recognition, activation and effector pathways. To allow in depth research on their modes of immune evasion, several mouse models for parasitic helminth infections have been established. Heligmosomoides polygyrus for example is a gastrointestinal nematode of rodents exhibiting a wide spectrum of immunomodulatory effects, mediated in part by soluble molecules released by adult worms in vitro, the excretory/secretory products (HES). HES is a potent inhibitor of dendritic cell (DC) activation by Toll-like receptor (TLR) ligands, completely abolishing LPS induced IL-12 production and reducing the upregulation of cell surface activation markers. As of now, neither the modulatory molecule nor its mechanism of action are known. Here, the effect of HES on TLR ligand induced DC maturation was characterized in considerably more detail compared to previous publications. It could be shown to inhibit DC maturation induced by various TLR ligands, on both protein and mRNA levels. These effects were comparable in both C57BL/6 and BALB/c derived cells; in contrast to this HES differentially affected alternative activation of BMDC from these two mouse strains. Although for most of the experiments GM-CSF differentiated BMDC were used, HES also inhibited LPS induced activation of splenic CD11c+ cells as well as the activation of all three populations described in Flt3-L differentiated BMDC - pDCs, CD11b+ cDCs and CD24+ cDCs. Furthermore, it could be shown here that HES also inhibits LPS induced maturation in human monocyte derived DCs. In the search for the component in HES responsible for its inhibition of TLR ligand induced DC maturation, exosome depleted HES rather than exosomes was inhibitory, and the effect was heat labile. This lead to the conclusion that the modulatory molecule has a protein component which is indispensable for its effect; following this reasoning HES was subjected to fractionation, with subsequent analysis of the fraction protein contents by mass spectrometry. The top nine candidate proteins were expressed recombinantly; however, the recombinants were not able to inhibit LPS induced DC activation. In parallel, experiments to elucidate the mechanism by which HES inhibits TLR ligand induced DC maturation were performed. This led to the conclusion that HES induces changes in the cells that, while not affecting the induction of signalling downstream of TLRs, do impair its maintenance. As a complement to these experiments, the transcriptomes of LPS and LPS+HES treated cells eight hours after LPS stimulation were compared. This revealed that transcripts encoding a number of transcription factors inducing the expression of activation markers after TLR ligation were reduced upon treatment of cells with HES, as were the transcript levels of IRAK2, a kinase necessary for persistent signalling. In addition, HES increased the transcript levels for several factors known to negatively regulate DC maturation, including ATF3. Furthermore, this analysis revealed changes in transcript levels of factors like HIF-1a, indicating an even greater reliance on aerobic glycolysis if cells were treated with HES, in addition to hints at increased ER and oxidative stress. In conclusion, this work narrows down the list of potential DC modulators in HES, gives a first insight into changes in DC metabolism induced by HES and sheds light on the role of a number of signalling pathways with important roles in DC activation as targets of DC inhibition by HES.

APA, Harvard, Vancouver, ISO, and other styles

9

Job, Christy Amelia. "The biogenesis of regulated secretory organelles." Thesis, University College London (University of London), 1995. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.309240.

Full text

APA, Harvard, Vancouver, ISO, and other styles

10

Alkhalife, Ibrahim S. M. "Axenic culture, infectivity and secretory proteins of Leishmania major." Thesis, University of Liverpool, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.366507.

Full text

APA, Harvard, Vancouver, ISO, and other styles

11

Rayner, Julian Charles. "Sorting of membrane proteins in the yeast secretory pathway." Thesis, University of Cambridge, 1997. https://www.repository.cam.ac.uk/handle/1810/284365.

Full text

APA, Harvard, Vancouver, ISO, and other styles

12

Gaudrey, Claire Anne. "Tissue transglutaminase : a new secretory protein." Thesis, Nottingham Trent University, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.245102.

Full text

APA, Harvard, Vancouver, ISO, and other styles

13

Oakley, Jacqueline D. "Interactions between proteins involved in mammalian endocytic and secretory pathways." Thesis, University of Bristol, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.404293.

Full text

APA, Harvard, Vancouver, ISO, and other styles

14

Johnson, Nicholas. "Post-translational translocation of secretory proteins at the endoplasmic reticulum." Thesis, University of Manchester, 2012. https://www.research.manchester.ac.uk/portal/en/theses/posttranslational-translocation-of-secretory-proteins-at-the-endoplasmic-reticulum(932aad49-0bb1-4fb2-a59f-def0112301be).html.

Full text

Abstract:

In eukaryotes, many secretory protein precursors are thought to translocate across the membrane of the endoplasmic reticulum (ER) co-translationally, in a process that is also dependent on their targeting via the signal recognition particle (SRP). However, a previously unsuspected number of metazoan secretory proteins have been shown to utilise a distinct, post-translational, mechanism(s) for their delivery to and translocation across the ER. Such proteins are unusually short; therefore SRP does not have sufficient opportunity to engage with their N-terminal signal sequence before polypeptide synthesis is complete. In this study, I examine the properties of secretory proteins that are known or potential candidates for post-translational translocation, and identify cytosolic components that mediate this process (Chapters 2.1 to 2.3). In vitro analysis using chimeric proteins and inhibitors of ER translocation suggested that both the signal sequence and mature domains of secretory proteins might influence their capacity for post-translational translocation (Chapter 2.1). These effects are most likely exerted by the Sec61 translocon of the ER membrane. The ER delivery of this class of proteins bears a striking resemblance to that of tail-anchored membrane proteins, with clear evidence for multiple overlapping pathways. Furthermore, a known tail-anchor protein delivery factor, TRC40, was found to mediate one of at least two distinct pathways that target short secretory proteins to the ER where they utilise the Sec61 complex for translocation (Chapter 2.2). During an in vitro screen of other cytosolic components potentially influencing post-translational translocation, Pex19 was observed to affect this process. Preliminary findings suggest that Pex19 appears to interact with proteins delivered to the ER post-translationally, and these observations merit further investigation (Chapter 2.3). Based on these results and current knowledge in the wider field, a working model describing the molecular mechanisms that underlie the post-translational translocation of short secretory proteins is presented, and I speculate on the delivery pathways and specific properties of these unique polypeptide substrates.

APA, Harvard, Vancouver, ISO, and other styles

15

Miranda, Kevin Charles. "Post-Golgi trafficking in the mammalian secretory pathway /." [St. Lucia, Qld], 2004. http://www.library.uq.edu.au/pdfserve.php?image=thesisabs/absthe18194.pdf.

Full text

APA, Harvard, Vancouver, ISO, and other styles

16

Kupzig, Sabine. "Identification and characterisation of two novel proteins of the secretory pathway." Thesis, University of Bristol, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.245577.

Full text

APA, Harvard, Vancouver, ISO, and other styles

17

Glenn, Daphne Elizabeth. "The role of GTP-binding proteins in regulated exocytosis." Thesis, University of Liverpool, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.367081.

Full text

APA, Harvard, Vancouver, ISO, and other styles

18

Gushue, Jennifer Nicole. "Characterization and regulation of the early secretory pathway by the p24 proteins." Thesis, McGill University, 2002. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=82887.

Full text

Abstract:

The secretory pathway of dog pancreas, liver parenchyma, cultured CHO cells and yeast has been analyzed with reagents focusing on the p24 family of membrane proteins. The results show that at least one of the p24s is ribosome associated in a nascent chain dependent manner. The cell free reconstitution of the endoplasmic reticulum (ER) cargo exit sites has demonstrated that the p24s are involved in the generation of both the cargo exit sites and vesicular tubular clusters (VTCs) in a COPI dependent manner. Work done in situ and in isolated subcellular fractions has indicated that the p24-enriched cis Golgi compartment is the primary site of retrieval of Golgi resident enzymes during membrane maturation as well as being the boundary for the major site of cargo (albumin) concentration. The morphogenic properties of the p24s were studied in yeast, with DNA chip analysis revealing 160 genes whose expression was altered coincident with the perturbation of members of the p24 family. Over expression of one of the members of the p24 family has led to the increase in ER membranes and a p24 enriched VTC compartment. Taken together, these data indicate that the p24s are present both when newly synthesized cargo enter and exit the ER and that the p24s are involved in the morphological transitions of the early secretory pathway.

APA, Harvard, Vancouver, ISO, and other styles

19

Li, Ling. "Effects of pancreatic secretory stress proteins (SSP) on human pancreatic stellate cells (hPSC)." [S.l. : s.n.], 2007. http://nbn-resolving.de/urn:nbn:de:bsz:289-vts-58568.

Full text

APA, Harvard, Vancouver, ISO, and other styles

20

Gleave, Terrence Lee. "KIM-2 : a model mammary epithelial cell line for the study of exocytosis." Thesis, University of Liverpool, 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.343755.

Full text

APA, Harvard, Vancouver, ISO, and other styles

21

Wetherell, Mark Anthony. "Individual differences in secretory immunoglobulin A (S-IgA) reactivity to acute stress." Thesis, University of Plymouth, 2002. http://hdl.handle.net/10026.1/2546.

Full text

Abstract:

Secretory immunoglobulin-A (S-IgA) is an antibody found on all surfaces of the common mucosa and serves as a first line of defence against pathogens. S-IgA is the predominant antibody in human secretions and unlike many other immune parameters, can be measured non-invasively in saliva. In addition to being an efficient indicator of health status, S-IgA levels are sensitive to variations in subjective and objective levels of stress, both of which are also influenced by state and trait factors. Stress is known to play an important role in susceptibility to infections of the common mucosa, and as such, the role of S-IgA as a potential moderating variable between stress and health is of increasing clinical importance. This thesis assessed the roles of retrospectively reported health status (minor health complaints) and state and trait factors upon levels of S-IgA following acute stress (S-IgA reactivity). Stress was manipulated using a multi-tasking performance battery, which unlike other laboratory based stressors is analogous to a variety of working environments. In a series of studies (3), S-IgA reactivity was observed following the stressor on one occasion, two occasions (24 hours apart) and following repeated stress on one occasion (cumulative acute stress). Volunteers classified as in poor health using a specifically designed health questionnaire, demonstrated consistently reduced S-IgA reactivity when compared to volunteers classified as being in good health. Furthermore, the discrepancy in S-IgA reactivity between good and poor health volunteers was most evident following cumulative acute stress. That is, poor health volunteers demonstrated progressive reductions in S-IgA reactivity as the accumulation of stress became greater. Volunteers in poor health were also characterised by negative state and trait characteristics, which in addition, were also independently associated with reduced S-IgA reactivity to acute stress. The findings indicate that negative state and trait characteristics are associated with reduced S-IgA reactivity to acute stress, levels of which influence post-stress susceptibility to illness. Further, deleterious effects of acute stress are most apparent in poor health volunteers following cumulative acute stress analogous with the stressors encountered in a variety of working environments.

APA, Harvard, Vancouver, ISO, and other styles

22

Li, Yongling. "Immunofluorescence investigations on neuroendocrine secretory protein 55 (NESP55) in nervous tissues /." Göteborg : Dept. of Medical Chemistry and Cell Biology, Institute of Biomedicine, Sahlgrenska Academy, Göteborg University, 2008. http://hdl.handle.net/2077/9892.

Full text

APA, Harvard, Vancouver, ISO, and other styles

23

Wang, Pengwei. "KMS, a group of novel plant endoplasmic reticulum proteins involved in the early secretory pathway." Thesis, Oxford Brookes University, 2010. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.515239.

Full text

APA, Harvard, Vancouver, ISO, and other styles

24

Parish, Lindsay A. "Protein trafficking and 4.1R relocalization in Plasmodium falciparum-infected erythrocytes." Thesis, Birmingham, Ala. : University of Alabama at Birmingham, 2009. https://www.mhsl.uab.edu/dt/2009p/parish.pdf.

Full text

APA, Harvard, Vancouver, ISO, and other styles

25

Szul, Tomasz J. "The role of GBF1 in Golgi biogenesis and secretory traffic." Thesis, Birmingham, Ala. : University of Alabama at Birmingham, 2009. https://www.mhsl.uab.edu/dt/2009p/szul.pdf.

Full text

APA, Harvard, Vancouver, ISO, and other styles

26

Denzel, Angela. "Generation and analysis of p23 and calnexin deficient mice." Thesis, University College London (University of London), 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.312166.

Full text

APA, Harvard, Vancouver, ISO, and other styles

27

Winkler, Sandra, Madlen Hempel, Sandra Brückner, Hans-Michael Tautenhahn, Roland Kaufmann, and Bruno Christ. "Identification of pathways in liver repair potentially targeted by secretory proteins from human mesenchymal stem cells." Universitätsbibliothek Leipzig, 2016. http://nbn-resolving.de/urn:nbn:de:bsz:15-qucosa-207430.

Full text

Abstract:

Background: The beneficial impact of mesenchymal stem cells (MSC) on both acute and chronic liver diseases has been confirmed, although the molecular mechanisms behind it remain elusive. We aim to identify factors secreted by undifferentiated and hepatocytic differentiated MSC in vitro in order to delineate liver repair pathways potentially targeted by MSC. Methods: Secreted factors were determined by protein arrays and related pathways identified by biomathematical analyses. Results: MSC from adipose tissue and bone marrow expressed a similar pattern of surface markers. After hepatocytic differentiation, CD54 (intercellular adhesion molecule 1, ICAM-1) increased and CD166 (activated leukocyte cell adhesion molecule, ALCAM) decreased. MSC secreted different factors before and after differentiation. These comprised cytokines involved in innate immunity and growth factors regulating liver regeneration. Pathway analysis revealed cytokine-cytokine receptor interactions, chemokine signalling pathways, the complement and coagulation cascades as well as the Januskinase-signal transducers and activators of transcription (JAK-STAT) and nucleotide-binding oligomerization domain-like receptor (NOD-like receptor) signalling pathways as relevant networks. Relationships to transforming growth factor beta(TGF-beta) and hypoxia-inducible factor 1-alpha (HIF1-alpha) signalling seemed also relevant. Conclusion: MSC secreted proteins, which differed depending on cell source and degree of differentiation. The factors might address inflammatory and growth factor pathways as well as chemo-attraction and innate immunity. Since these are prone to dysregulation in most liver diseases, MSC release hepatotropic factors, potentially supporting liver regeneration.

APA, Harvard, Vancouver, ISO, and other styles

28

Dahm, Christina Catherine. "S-nitrosothiol formation in mitochondrial proteins." Thesis, University of Cambridge, 2005. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.615098.

Full text

APA, Harvard, Vancouver, ISO, and other styles

29

Stiles, Bangyan Li. "Keratinocyte secretory phospholipase A₂s : its characterization, modulation, and role in mouse skin carcinogenesis /." Digital version accessible at:, 1998. http://wwwlib.umi.com/cr/utexas/main.

Full text

APA, Harvard, Vancouver, ISO, and other styles

30

Muckley, Philippa L. "A study of RAB3 GTP-binding proteins in the secretory pathways of a mouse pituitary cell line." Thesis, Oxford Brookes University, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.287728.

Full text

APA, Harvard, Vancouver, ISO, and other styles

31

Schlegel, Susan. "From protein production to genome evolution in Escherichia coli." Doctoral thesis, Stockholms universitet, Institutionen för biokemi och biofysik, 2013. http://urn.kb.se/resolve?urn=urn:nbn:se:su:diva-94993.

Full text

Abstract:

The aim of my Ph.D. studies was to improve production yields of membrane- and secretory proteins in the widely used E. coli protein production strain BL21(DE3). In this strain expression of the gene encoding the protein of interest is driven by the powerful T7 RNA polymerase (T7 RNAP) whose gene is located on the chromosome and under control of the strong, IPTG-inducible lacUV5 promoter. Unfortunately, the production of many membrane and secretory proteins is 'toxic' to BL21(DE3), resulting in poor growth and low production yields. To understand this ‘toxicity’, the BL21(DE3) derived mutant strains C41(DE3) and C43(DE3) were characterized. Somehow, these strains can efficiently produce many ‘toxic’ membrane and secretory proteins. We showed that mutations weakening the lacUV5 promoter are responsible for this. These mutations result in a slower onset of protein production upon the addition of IPTG, which avoids saturating the Sec-translocon capacity. The Sec-translocon is a protein-conducting channel in the cytoplasmic membrane mediating the biogenesis of membrane proteins and translocation of secretory proteins. Next, we constructed a BL21(DE3)-derivative, Lemo21(DE3), in which the activity of T7 RNAP can be precisely controlled by titrating in its natural inhibitor T7 lysozyme using the rhamnose promoter system. In Lemo21(DE3), the expression level of genes encoding membrane and secretory proteins can be set such that the Sec-translocon capacity is not saturated. This is key to optimizing membrane and secretory protein production yields. Finally, reconstructing the evolution of C41(DE3) from BL21(DE3) in real time showed that during its isolation C41(DE3) had acquired mutations critical for surviving the starvation conditions used, and provided insight in how the mutations in the lacUV5 promoter had occurred.

At the time of the doctoral defense, the following paper was unpublished and had a status as follows: Paper 3: Manuscript.

APA, Harvard, Vancouver, ISO, and other styles

32

Ackroyd, Mark. "Development of model systems to study the function of excretory-secretory proteins from the parasitic nematode 'Trichinella spiralis'." Thesis, University of Aberdeen, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.430389.

Full text

Abstract:

Invasion of host muscle by the larvae of the nematode parasite Trichinella spiralis is associated with a series of ultrastructural, biochemical and molecular changes to the host muscle fibre that amalgamates in the formation of a unique host-parasite complex. It is hypothesised that proteins that are secreted or excreted (ES proteins) by the muscle larva are responsible for triggering some or all of the changes to the host cell. No such effector molecules have yet been identified although a number of candidate molecules are now emerging. There is therefore a need for an experimental system (or systems) that will enable the study of function of candidate ES proteins as they arise. In this investigation I describe the development of three model systems that allow the characterisation of different aspects of putative protein function with the aim of determining whether a candidate protein could contribute to the formation of the muscle host parasite complex. These models included: firstly, a cell culture model to investigate the effect of T. spiralis ES proteins of the differentiation of the myogenic cell line C2C 12; secondly, two in vivo heterologous expression systems to investigate the secretion and localisation of T. spiralis ES proteins; and thirdly, a cell-free in vitro translation system to investigate the secretion and potential processing and post-translation modifications of candidate T. spiralis ES proteins. The model systems described have enabled conclusions to be drawn on the potential role that a candidate ES protein, TsJ5, may play in both the biology of the muscle larvae and within the parasitised host cell.

APA, Harvard, Vancouver, ISO, and other styles

33

Bian, Shumin. "Fe-S proteins : cluster assembly and degradation /." The Ohio State University, 1998. http://rave.ohiolink.edu/etdc/view?acc_num=osu1487952208109007.

Full text

APA, Harvard, Vancouver, ISO, and other styles

34

DiLuna, Francis Anthony. "INHIBITORY PROPERTIES OF MICROPLITIS CROCEIPES TERATOCYTE SECRETORY PRODUCTS AND THE RECOMBINANT PROTEIN TSP14 ON PROTEIN SYNTHESIS." Lexington, Ky. : [University of Kentucky Libraries], 2003. http://lib.uky.edu/ETD/ukyento2003t00126/FADTHES.pdf.

Full text

Abstract:

Thesis (M.S.)--University of Kentucky, 2003.
Title from document title page (viewed June 21, 2004). Document formatted into pages; contains xii, 122 p. : Ill. Includes abstract and vita. Includes bibliographical references (p. 106-121).

APA, Harvard, Vancouver, ISO, and other styles

35

Braganza, Annabel M. H. (Mary Helen). "Characterization of nascent enamel proteins translated in vitro from mRNA specific for the secretory and maturation stages of amelogenesis." Thesis, McGill University, 1995. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=22851.

Full text

Abstract:

Enamel proteins are extracellular matrix proteins expressed throughout enamel formation. However, questions concerning their numbers and origins are still somewhat ambiguous. To characterize the nascent enamel proteins, cell free translation and immunoprecipitation procedures were performed using poly(A)$ sp{+}$RNA isolated from freeze-dried segments of rat incisor enamel organs at the secretory, early maturation, and mid maturation stages of amelogenesis. Phosphoimaging revealed that the enamel organ produces enamel proteins continuously throughout enamel development albeit in decreasing number and intensity. Ten enamel proteins were translated at the secretory stage and ranged in molecular weight from 80 to 18 kDa, resembling the number (Simmer et al., 1994) and the size (DenBesten et al., 1992) of RNA transcripts recently described for murine and rat. At the early and mid maturation stages, the enamel proteins span a 27 to 68 kDa region. This selected expression of enamel proteins at each stage of development suggests that specific proteins may be important for the maturation process.

APA, Harvard, Vancouver, ISO, and other styles

36

Shaw, Amanda Marie. "Interactions of the growth hormone secretory axis and the central melanocortin system." [Gainesville, Fla.] : University of Florida, 2004. http://purl.fcla.edu/fcla/etd/UFE0008380.

Full text

Abstract:

Thesis (Ph.D.)--University of Florida, 2004.
Typescript. Title from title page of source document. Document formatted into pages; contains 142 pages. Includes Vita. Includes bibliographical references.

APA, Harvard, Vancouver, ISO, and other styles

37

Roeske, Cassandra. "Role of the Heterotrimeric Go Protein Alpha-subunit on the Cardiac Secretory Phenotype." Thèse, Université d'Ottawa / University of Ottawa, 2013. http://hdl.handle.net/10393/24191.

Full text

Abstract:

Atrial natriuretic factor (ANF) is a polypeptide hormone produced in heart atria, stored in atrial secretory granules and released into the circulation in response to various stimuli. Proper sorting of ANF at the level of the trans-Golgi network (TGN) is required for the storage of ANF in these specific granules, and this sorting of hormones has been found to be associated with G-proteins. Specifically, the Go protein alpha-subunit (Gαo) was established to participate in the stretch-secretion coupling of ANF, but may also be involved in the transporting of ANF from the TGN into atrial granules for storage and maturation. Based on knowledge of Gαo involvement in hormone production in other endocrine tissues, protein-protein interactions of Gαo and proANF and their immunochemical co-localization in granules, the direct involvement of these two proteins in atrial granule biogenesis is probable. In this study, mice were created using the Cre/lox recombination system with a conditional Gαo knockout in cardiocytes to study and characterize ANF production, secretion and granule formation. Deletion of this gene was successful following standard breeding protocols. Characterization and validation of cellular and molecular content of the knockout mice through mRNA levels, protein expression, peptide content, electron microscopy, and electrocardiography determined that a significant phenotypic difference was observed in the abundance of atrial granules. However, Gαo knockout mice did not significantly alter the production and secretion of ANF and only partially prevented granule biogenesis, likely due to incomplete Gαo knockout. These studies demonstrate an involvement of Gαo in specific atrial granule formation.

APA, Harvard, Vancouver, ISO, and other styles

38

Herrmann, Lydia, Caspar Wiegmann, Annika Arsalan-Werner, Isabel Hilbrich, Carsten Jäger, Katharina Flach, Anne Suttkus, Ingolf Lachmann, Thomas Arendt, and Max Holzer. "Hook proteins." Universitätsbibliothek Leipzig, 2015. http://nbn-resolving.de/urn:nbn:de:bsz:15-qucosa-169817.

Full text

Abstract:

Defects in intracellular transport are implicated in the pathogenesis of Alzheimer’s disease (AD). Hook proteins are a family of cytoplasmic linker proteins that participate in endosomal transport. In this study we show that Hook1 and Hook3 are expressed in neurons while Hook2 is predominantly expressed in astrocytes. Furthermore, Hook proteins are associated with pathological hallmarks in AD; Hook1 and Hook3 are localized to tau aggregates and Hook2 to glial components within amyloid plaques. Additionally, the expression of Hook3 is reduced in AD. Modelling of Hook3 deficiency in cultured cells leads to slowing of endosomal transport and increases β-amyloid production. We propose that Hook3 plays a role in pathogenic events exacerbating AD.

APA, Harvard, Vancouver, ISO, and other styles

39

Arthur, Ian D. "The association between the major endometrial secretory proteins (IGFBP-1 and PP14) and the reproductive response in assisted conception cycles." Thesis, University of Southampton, 1995. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.296243.

Full text

APA, Harvard, Vancouver, ISO, and other styles

40

Boyes, Barry Edward. "An immunochemical and immunocytochemical study of the S-100b protein." Thesis, University of British Columbia, 1985. http://hdl.handle.net/2429/24485.

Full text

Abstract:

This thesis describes an immunochemical and immunocytochemical study of the bovine brain S-lOOb protein. The two major forms of the S-100 isoproteins (S-lOOa and S-lOOb) were purified to apparent homogeneity from bovine brain. A polyclonal rabbit antiserum to the S-lOOb protein was prepared. The antiserum was characterized by solid phase immunochemical methods. The S-lOOb derived antiserum displayed a high degree of specificity for S-lOOb, but also crossreacted with the purified S-lOOa protein. The characteristics of the immunochemical reactivity of the antiserum towards these two isoproteins suggests the antiserum has specificity for the 6-subunit of the S-100 proteins. An immunohistochemical analysis of the cellular localization of S-lOOb immunoreactivity was undertaken. In the adult rat brain only the astrocytes were S-lOOb immunoreactive. This conclusion is supported by the morphological characteristics of the immunolabelled cells, as well as the observed co-localization of the immunoreactivities for S-lOOb and the Glial Fibrillary Acidic protein (GFAP), the major protein of the astrocyte intermediate filaments. These two antigens were always found to coexist. The immunolabelling of rat brain astrocytes by the S-lOOb derived antiserum stained the entire cell, yielding more complete morphological detail than is possible with the GFAP immunohistochemistry, which only labels the filamentous glial processes. It is concluded that S-lOOb immunohistochemistry could be of general utility in the investigation of astrocyte morphology. The present results also determine that the as yet unknown biological function(s) of the S-100 proteins must be related to a property of astrocytes.
Medicine, Faculty of
Graduate

APA, Harvard, Vancouver, ISO, and other styles

41

Li, Jianhui. "Cornichon Proteins: Unexpected Roles in Plant Pathogen Infection, ER Morphology Maintenance and Pollen Development." Diss., Virginia Tech, 2017. http://hdl.handle.net/10919/77687.

Full text

Abstract:

Cornichon (CNI) proteins are a conserved family of proteins among eukaryotes, from Erv14 in the yeast Saccharomyces cerevisiae to CNI homologs (CNIHs) in mammals and plants. Erv14 functions as a cargo receptor of coat protein complex II (COPII) for protein trafficking from the endoplasmic reticulum (ER) to the Golgi apparatus, en route to their final destinations. By interacting with specific cargo proteins, CNI proteins regulate key steps of embryo polarity in Drosophila, budding in yeast, and synaptic transmission in the mammalian brain. However, we have very limited understanding of plant CNIHs. Positive-strand RNA viruses assemble their viral replication complexes (VRCs) at specific host organelle membranes. With a better understanding of host factors involved in targeting viral replication proteins to the preferred organelles, we expect to block trafficking of viral replication proteins and thus, viral infection, by manipulating the required host proteins. Brome mosaic virus (BMV) is a model of positive-strand RNA viruses and its replication can be recapitulated in yeast. Importantly, BMV replication protein 1a is the only required viral protein to form VRCs at the perinuclear ER membrane in yeast. I demonstrate that Erv14 and COPII coat proteins are required for targeting BMV 1a to the perinuclear ER in yeast, suggesting a novel function of COPII vesicles in protein trafficking to the perinuclear ER membrane and in the BMV VRC formation. As for cellular functions, I show that plant CNIHs complement the defective distribution of BMV 1a in yeast mutant lacking Erv14. Taking advantage of Arabidopsis thaliana knockout mutants and knockdown of gene expression in Nicotiana benthamina, I also discover that CNIHs unexpectedly play crucial roles in pollen development, infection of a bacterial pathogen, and maintenance of ER tubules. I further confirm that CNI proteins are also required for maintaining ER tubules in yeast, suggesting a novel and conserved role in shaping ER morphology. Therefore, these findings indicate the functional diversity and redundancy of CNI proteins in key cellular processes and suggest a novel strategy to control plant pathogenic viruses and bacteria by manipulating plant CNIHs.
Ph. D.

APA, Harvard, Vancouver, ISO, and other styles

42

Smit, Egbert. "Lactobacillus S-layer proteins structure-function relationship and application potential /." [S.l. : Amsterdam : s.n.] ; Universiteit van Amsterdam [Host], 2002. http://dare.uva.nl/document/63739.

Full text

APA, Harvard, Vancouver, ISO, and other styles

43

Rees, Christopher Gareth. "Identification and characterisation of heparin binding proteins in S. Aureus." Thesis, University of Liverpool, 2009. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.526803.

Full text

APA, Harvard, Vancouver, ISO, and other styles

44

Dutta, Chaitali. "Checkpoint Regulation of S-Phase Transcription: A Dissertation." eScholarship@UMMS, 2008. https://escholarship.umassmed.edu/gsbs_diss/391.

Full text

Abstract:

The DNA replication checkpoint transcriptionally up-regulates genes that allow cells to adapt to and survive replication stress. Our results show that, in the fission yeast Schizosaccharomyces pombe, the replication checkpoint regulates the entire G1/S transcriptional program by directly regulating MBF (aka DSC1), the G1/S transcription factor. Instead of initiating a checkpoint-specific transcriptional program, the replication checkpoint targets MBF to maintain the normal G1/S transcriptional program during replication stress. We propose a mechanism for this regulation, based on in vitrophosphorylation of the Cdc10 subunit of MBF by the Cds1 replication-checkpoint kinase. Substitution of two potential phosphorylation sites with phospho-mimetic amino acids suffice to promote the checkpoint transcriptional program, suggesting that Cds1 phosphorylation directly regulates MBF-dependent transcription. The conservation of MBF between fission and budding yeast, and recent results implicating MBF as a target of the budding yeast replication checkpoint, suggest that checkpoint regulation of the MBF transcription factor may be a conserved strategy for coping with replication stress. Furthermore, the structural and regulatory similarity between MBF and E2F, the metazoan G1/S transcription factor, suggests that this checkpoint mechanism may be broadly conserved among eukaryotes. Our result shows that both the replication checkpoint and the S-phase DNA damage checkpoint are involved in activating MBF regulated S-phase gene transcription and that this coordinated transcriptional response is beneficial for survival during replication stress. I demonstrate that the beneficial role of the transcriptional response during checkpoint activation is mediated by three major MBF transcripts: cdc22, mrc1 and mik1. Mrc1 dependent stabilization of stalled fork is important during S phase arrest. However, cells ability to prevent mitosis (Mik1 dependent) along with stable fork (Mrc1 dependent) both are crucial for survival. Our data also suggest that the level of Cdc22 is a determining factor for replication checkpoint activation and when over-expressed can alleviate the effects not only in HU but also in MMS.

APA, Harvard, Vancouver, ISO, and other styles

45

Moustafa, Amr [Verfasser], and Christian [Akademischer Betreuer] Betzel. "Structural and Functional Analyses of Secretory and Excretory Proteins from Onchocerca volvulus as Basis for Rational Drug Design / Amr Moustafa. Betreuer: Christian Betzel." Hamburg : Staats- und Universitätsbibliothek Hamburg, 2016. http://d-nb.info/1098428927/34.

Full text

APA, Harvard, Vancouver, ISO, and other styles

46

Moustafa, Amr Verfasser], and Christian [Akademischer Betreuer] [Betzel. "Structural and Functional Analyses of Secretory and Excretory Proteins from Onchocerca volvulus as Basis for Rational Drug Design / Amr Moustafa. Betreuer: Christian Betzel." Hamburg : Staats- und Universitätsbibliothek Hamburg, 2016. http://nbn-resolving.de/urn:nbn:de:gbv:18-78577.

Full text

APA, Harvard, Vancouver, ISO, and other styles

47

Bauer, Roslyn A. "Characterization of sorting motifs in the dense core vesicle membrane protein phogrin /." Connect to full text via ProQuest. Limited to UCD Anschutz Medical Campus, 2008.

Find full text

Abstract:

Thesis (Ph.D. in Cell Biology, Stem Cells, & Development) -- University of Colorado Denver, 2008.
Typescript. Includes bibliographical references (leaves 138-155). Free to UCD Anschutz Medical Campus. Online version available via ProQuest Digital Dissertations;

APA, Harvard, Vancouver, ISO, and other styles

48

Grognono-Thomas, Rosemary. "The role of (S)-layer proteins in ovine Campylobacter fetus infections." Thesis, Imperial College London, 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.312325.

Full text

APA, Harvard, Vancouver, ISO, and other styles

49

Pakdel, Mehrshad [Verfasser], and Reinhard [Akademischer Betreuer] Fässler. "Activity of the SPCA1 calcium ATPase couples sphingomyelin synthesis to sorting of secretory proteins in the trans-Golgi network / Mehrshad Pakdel ; Betreuer: Reinhard Fässler." München : Universitätsbibliothek der Ludwig-Maximilians-Universität, 2019. http://d-nb.info/1194835341/34.

Full text

APA, Harvard, Vancouver, ISO, and other styles

50

Learman, Sarah Sebring. "Small-Molecule Control of Kinesin-5 Proteins." Diss., Virginia Tech, 2008. http://hdl.handle.net/10919/26579.

Full text

Abstract:

Mitosis, or cell division, is the mechanism by which cells divide and is an intricate process requiring the action and control of numerous proteins. Such proteins serve either as structural entities within the mitotic spindle, or perform the â workâ within the apparatus. In particular, Kinesin-5 motor proteins, a subset within the kinesin motor protein superfamily, are primarily responsible for organization of microtubules (MTs) within the mitotic apparatus, and are consequently vital for efficient mitosis. These proteins utilize energy from ATP hydrolysis in order to â walkâ along antiparallel MTs, positioning them into the bipolar mitotic spindle. Loss of Kinesin-5 activity results in formation of a monoastral spindle and subsequent cell cycle arrest. Recently, a wide variety of small molecules have been identified that possess the ability to inhibit certain Kinesin-5 motors. Such compounds, including monastrol (the first Kinesin-5 inhibitor identified), have been employed to study Kinesin-5 activity. A thorough understanding of Kinesin-5 function, combined with the ability to specifically target these proteins with small molecules, may provide the capability to control cell division and may therefore have significant implications in anti-cancer therapies. The following dissertation describes research that utilizes small molecules to probe the function (ATPase activity and MT interactions) of various Kinesin-5 proteins and provides information that will lead to a better understanding of exactly how such proteins function in vivo. Further, a greater knowledge of Kinesin-5 protein activity as well as specific interactions with small-molecule compounds, may lead to the development of more potent, less toxic anti-cancer drugs.
Ph. D.

APA, Harvard, Vancouver, ISO, and other styles

Dissertations / Theses on the topic 'Secretory (E/S) proteins'

Create a spot-on reference in APA, MLA, Chicago, Harvard, and other styles