Rozprawy doktorskie: „Receptor affinity”

1

Scammells, Peter J., i n/a. "Pyrazolo(3,4-d)Pyrimidines and adenosine receptors: a structure/activity study". Griffith University. Division of Science and Technology, 1990. http://www4.gu.edu.au:8080/adt-root/public/adt-QGU20050826.141630.

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Pyrazolopyrimidines are a general class of compounds which exhibit Aj adenosine receptor affmity. A number of pyrazolo(3,4-d)pyrimidine analogues of isoguanosine and i-methylisoguanosine has been synthesised. All compounds were tested forAi adenosine receptor affinity using a (311) R-PIA competitive binding assay. The N-i and N-5 positions were substituted with a number of different ailcyl and aryi groups. 3-Chiorophenyl substitution of the N-i position and butyl substitution of the N-5 position greatly enhanced the overall adenosine receptor affinity. Substitution by a methyl group at the N-7 position fixed the C-4 position in the imino tautomeric form. This resulted in a marked reduction in activity. The substitution of the N-2 position with a phenyl group produced an analogue with a similar structure to i,3-dipropyl-8-(2-amino-4-chlorophenyl)xanthine (PACPX). A 2-phenyl substituent was favourable for interaction with the adenosine receptor. A number of pyrazolo(3,4-d)pyrirnidine analogues of 4,6-bis-a-carbamoylethylthio-i-phenylthiopyrazolo(3,4-d)pyrinhidine (DJB-KK) has also been synthesised and tested for Aj adenosine receptor affinity. 4,6-Bis-alkylthio-1-phenylpyrazolo(3,4-d)pyrimidines with a-carbamoylethyl and u-carbamoylpropyi groups were compared. The additional methyiene of the a-carbamoylpropyl group produced increased adenosine receptor affinity. 6-a-Carbamoylethylthio-4-mercapto-1-phenylpyrazolo(3,4-d)pyrimidine and 4-cc-carbamoylethylthio- i-phenylpyrazolo(3,4-dlpyrimidine were compared. Substitution of the C-6 position maintained activity, while substitution of the C-4 reduced activity.

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2

Jackson, Leila J. "The dynamic regulation of the low affinity IGE receptor by toll like receptor and B cell receptor agonists /". Connect to full text via ProQuest. Limited to UCD Anschutz Medical Campus, 2008.

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Thesis (Ph.D. in Immunology) -- University of Colorado Denver, 2008.
Typescript. Includes bibliographical references (leaves 122-129). Free to UCD Anschutz Medical Campus. Online version available via ProQuest Digital Dissertations;

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3

Sundaramoorthy, Meena. "MODULATION OF HIGH AFFINITY HORMONE BINDING TO LH/CG RECEPTOR". UKnowledge, 2002. http://uknowledge.uky.edu/gradschool_theses/209.

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Precise control of physiological phenomena is performed by various kinds of receptormediated signaling. The vast majority of receptors belong to the superfamily of G proteincoupled receptors (GPCRs), which form one of the largest protein families. In theclassical model of GPCR signaling, stimulation of seven transmembrane spanning GPCRleads to the activation of heterotrimeric G proteins, which dissociate into a and bgsubunits. The subunits activate effector molecules, which include second messengergenerating systems, giving rise to various kinds of cellular responses. The LH/CGreceptor is a member of the glycoprotein hormone receptor family along with the FSHand TSH receptors, which belongs to the GPCR superfamily. Human chorionicgonadotropin (hCG) binds to the exodomain of LH/CG receptor and the resulting hCGexodomaincomplex is thought to interact with the endodomain of the receptor to bringabout hormone signal.Unfortunately, little evidence is available for the precise hormonecontact points in the exo domain and endo domain of the receptor. The affinity ofhormone binding to the exodomain was enhanced when the endodomain was truncated.This suggests that the endodomain modulates the hormone binding to the exodomain ofthe receptor. To understand this, the role of exoloop 2 on the modulation of high affinityhormone binding to the exodomain was studied using photoaffinity labeling technique.

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4

Smith, Lucy. "Investigating inhibitors of the IgE:high affinity receptor protein-protein interaction". Thesis, Imperial College London, 2014. http://hdl.handle.net/10044/1/44210.

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The protein-protein interaction (PPI) between immunoglobulin E (IgE) and its high affinity receptor (FcεRI) is an important part of the allergic response. Inhibition of the IgE:FcεRI interaction is a key strategy for the development of allergy treatments. This PPI has been validated as a therapeutic target by the humanised monoclonal antibody omalizumab, which binds to IgE and prevents the formation of the IgE:FcεRI complex and has proved successful at treating allergic asthma. However, small molecule inhibitors of the IgE:FcεRI PPI that are orally available would be a more desirable form of treatment. This thesis describes the design, synthesis and testing of two series of inhibitors of the IgE:FcεRI interaction; small molecules based on the natural product aspercyclide A and short, linear peptides based on a key binding epitope of FcεRI. It also describes the development of a high-throughput time resolved fluorescence resonance energy transfer (TR-FRET) assay to test inhibitors and subsequent X-ray crystallography and SPR experiments to further investigate the mode of action of the inhibitors. An analogue of aspercyclide A has shown inhibition of the IgE:FcεRI interaction in the micromolar range and an improved potency compared to the natural product itself. A number of 8-residue, linear peptides have been found to inhibit the IgE:FcεRI PPI in the micromolar range when tested in the TR-FRET assay. The most potent peptide has been biotinylated and immobilised for SPR experiments with IgE and FcεRI. These SPR experiments suggest that the peptide inhibits the IgE:FcεRI interaction by binding to the high affinity receptor rather than to IgE.

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5

Turner, Helen Cathryn. "Signal transduction by the high affinity receptor for immunoglobulin E, FcER1". Thesis, University College London (University of London), 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.301134.

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6

Mueller, Thomas. "Systems for measuring B cell receptor affinity maturation in germinal centres". Thesis, University of Birmingham, 2016. http://etheses.bham.ac.uk//id/eprint/7130/.

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Germinal Centres (GCs) play a central role in adaptive immunity; involving processes of cell migration, clonal expansion, hypermutation, and selection. To elucidate the role of affinity in regulating these processes, a technique for measuring B cell receptor affinity maturation in GCs in situ was developed. To facilitate interrogation of individual antibody-antigen interactions, atomic force microscopy (AFM) was chosen, offering nanometre positional resolution, and pico-Newton force sensitivity. Specificity of gold-coated AFM cantilevers towards the targeted receptors was achieved via a bespoke modification scheme, using self-assembled amine-terminated alkanethiol to facilitate attachment of the receptor specific antigen NP. Influences on molecule deposition and subsequent NP addition were investigated, as were control measures facilitating identification of successful modifications (Chapter 4). Effects of sample preparation techniques on AFM adhesion measurements were investigated (Chapter 5). Subsequently, the developed AFM technique was applied in interrogation of B cells and hybridomas – expressing receptors of varying affinity towards NP – and two GCs in tissue sections (Chapter 6). For the automated and unbiased evaluation of large amounts of varying AFM data, bespoke data analysis methods were developed. The project finds that AFM is capable of quantifying specific antibody-antigen interactions, but was unable to measure these in tissue sections. Possible reasons preventing such measurements are discussed.

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7

Timm, David Eugene. "Conformation, stability and interactions of the neurotrophins and the low-affinity neurotrophin receptor". Case Western Reserve University School of Graduate Studies / OhioLINK, 1993. http://rave.ohiolink.edu/etdc/view?acc_num=case1057179020.

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8

Wahlström, Niklas. "Synthesis of indoles, bisindoles and indolocarbazoles : high affinity aryl hydrocarbon receptor ligands /". Stockholm, 2004. http://diss.kib.ki.se/2004/91-7140-016-8/.

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9

梁頌偉 i Chung-wai Leung. "Novel gene transfer vector targeted high affinity IL-2 receptor bearing cell". Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2001. http://hub.hku.hk/bib/B31226280.

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10

Leung, Chung-wai. "Novel gene transfer vector targeted high affinity IL-2 receptor bearing cell /". Hong Kong : University of Hong Kong, 2001. http://sunzi.lib.hku.hk/hkuto/record.jsp?B25248698.

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11

Bruger, Annika Målin. "TCR signalling in response to affinity stimulation". Thesis, University of Oxford, 2013. http://ora.ox.ac.uk/objects/uuid:5f9c1001-6c43-472f-a495-c9573b54a84a.

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T cells are an essential part of the adaptive immune system and protect the body from intracellular infections. The specificity with which αßTCR-bearing T cells recognize cognate antigen presented on MHC molecules is paramount to maintaining the balance between mounting effector functions against pathogens and establishing peripheral tolerance to self. The mechanism by which T cells translate qualitative differences in TCR:pMHC binding to sensitive proximal signalling events which ultimately result in specific Tcell effector responses to infected cells but not to self is mostly unknown. To address how T cell signalling responds to qualitative differences in TCR triggering by pMHC, I established a system of stimulating T cells bearing the 1G4 TCR specifically in vitro with a panel of four NY-ESO-1_156-165 peptide variant MHC tetramers. Single amino acid substitutions to the NY-ESO-1_156-165 peptide conferred a maximum 35-fold difference in the monomeric affinity for the 1G4 TCR. The system allows the highly controlled investigation of very rapid TCR proximal signalling events simultaneously and quantitatively using flow cytometry. Stimulations with pMHC tetramers showed rapid sensitive sequential signalling responses which were able to confer ligand discrimination. Very early signalling events such as CD3ζ phosphorylation showed analogue responses to the different affinity pMHC tetramers. Later signalling events including phospho-ERK showed a distinct on/off switch-like response. The amplitude of the very early analogue signalling responses determined the extent of later digital ERK signals. This indicates that a certain analogue signalling threshold must be passed to result in T cell activation. The thymocyte protein Themis has been shown proximal TCR signalling to modulate thymocyte selection thresholds. Its deletion results in profound defects in positive thymocyte selection. Themis locates to the LAT signalosome of the TCR signalling cascade via Grb2, yet its molecular function is unknown. Employing the system I established, I demonstrate that Themis-k/d cells show increased levels of CD3z-chain phosphorylation, phospho-ERK signalling and signal-induced apoptosis which was independent of the ERK signal. This shows that Themis is a global attenuator of proximal TCR signalling. We are currently investigating possible associations of Themis to proteins phosphastases such as SHP-1 which could attenuate TCR proximal protein tyrosine signalling events.

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12

Noble, April R. "Synthesis of Amphibian Alkaloids and Synthesis and Affinity of Novel Cannabinoid Receptor Ligands". ScholarWorks@UNO, 2009. http://scholarworks.uno.edu/td/1103.

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Amphibian alkaloids are attractive targets for synthesis due to their biological activity. An important class of amphibian alkaloids is the 2,5-disubstituted pyrrolidine-based family of compounds. There are many synthetic approaches for the preparation of the trans-2,5- disubstituted pyrrolidines, but methods for the construction of the cis-2,5-pyrrolidines are limited. Therefore, it was desired to develop an enantioselective approach for the preparation of cis-2,5-disubsituted pyrrolidines. (+)-Tropin-2-one derived from cocaine was used as starting material to exploit the inherent stereochemistry for construction of the cis-pyrrolidine ring. This permitted the unequivocal assignment of the absolute configuration of the target pyrrolidine. The structurally simple pyrrolidine alkaloid, 225H, was selected as a target to develop a general synthetic approach. The enantioselective synthesis of 225H was achieved in nine steps and good overall yield. The search for potent cannabinoid receptor partial agonist ligands as potential marijuana addiction therapeutic agents has led to an investigation of the synthesis of diaryl ether hybrid analogues of BAY 59-3074. A series of 2-(3-alkyl-5-hydroxyphenoxy)-6- (trifluoromethyl)benzonitriles, 3-(2-cyano-3-(trifluoromethyl)phenoxy)phenylalkanoates, and (3- (benzyloxy)phenoxy)-6-(trifluoromethyl)benzonitriles were synthesized and evaluated in vitro for CB1 affinity. The olivetol diaryl ether analogue was the most potent ligand of the alkyl series, but the diaryl ester analogues exhibited modest affinity for CB1 receptors. The most potent compound of the series was the 2-(3-(benzyloxy)phenoxy)-6- (trifluoromethyl)benzonitrile.

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13

Li, Airong. "Variants of the #beta# subunit of the high affinity IgE receptor and atopy". Thesis, University of Oxford, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.318677.

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14

Kokalaki, E. K. "Affinity gradient of chimeric antigen receptor T-cells against low-antigen density target". Thesis, University College London (University of London), 2017. http://discovery.ucl.ac.uk/1563780/.

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Adoptive Cell Therapy (ACT) is a novel approach to cancer treatment, which implements the use of the patient T-cells redirected to eradicate tumour cells. The T-cells are engineered to express a Chimeric Antigen Receptor (CAR) that bestows specificity for an arbitrary antigen. CAR-T cell clinical trials have shown tumour eradication of leukaemia, but modest results against solid tumours. There is an ongoing debate regarding the correlation of CAR affinity and antigen-density recognition threshold, which we sought to investigate. Although the TCRs sensitivity against low antigen targets is superior at low-affinity, the effect of affinity in CAR efficacy and sensitivity is controversial. In order to examine the correlation between CAR efficacy and affinity, we designed a CAR against an intracellular protein in melanoma. Tyrosinase-Related Protein 1 (Tyrp1) resides in the surface of the organelles called melanosomes. However, Tyrp1 is also trafficked to the surface at a low density. We mutated an anti-Tyrp1 single-chain Variable Fragment (scFv) to acquire eight mutants of various affinities. The affinity range was 0.74nM-54.3nM. Including the wild-type scFv, these nine affinity gradient CARs were challenged with a cell line expressing low, medium and high densities of cell-surface Tyrp1. Three mutant scFv CARs were superior to the wild-type scFV. However, the affinities of those superior CARs was variant. There was no pattern observed to suggest a direct correlation between the affinity and the CAR efficacy. In contrast to previous publications, this study shows that affinity does not play a key role in determining the CAR efficacy. We hypothesise that the difference in CAR efficacy depends on a combination of factors, such as scFv stability and affinity, CAR density, and antigen density. However, the complex interaction of these parameters, as well as further confounding factors, renders the deduction of a pattern challenging. In order overcome this complexity, a multivariate analysis of all the parameters together is necessary.

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15

Dias, Ana Margarida Gonçalves Carvalho. "Development of an affinity pair “Tag-Receptor” for recombinant protein expression and purification". Master's thesis, Faculdade de Ciências e Tecnologia, 2010. http://hdl.handle.net/10362/10924.

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Dissertação apresentada para a obtenção do Grau de Mestre em Biotecnologia, pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia
The main objective of this work was the development of an affinity pair for the purification of recombinant proteins. In this work, ligands based on the Ugi Reaction and the 1,3,5-Triazine scaffold were synthesised in solid-phase and screened for binding to an affinity tag, an hexapeptide constituted by asparagine aminoacid (N). The ligands were tested against pure solutions of the hexapeptide and Green Fluorescence Protein (GFP), used as a model protein. The ligands that had the highest affinity for the hexapeptide and lowest affinity for the protein were selected for further studies with cellular extracts. The cellular extracts were produced in HEK 293T cells transfected with two designed vectors: one containing the GFP tagged with the affinity tag, and the other containing GFP without tag. The efficient expression of a recombinant GFP fused with the designed affinity tag was demonstrated. The cellular extracts were then loaded onto chromatographic columns containing the lead ligands immobilised onto agarose, and the amount of total protein and GFP bound and eluted noted. The results demonstrated that the Ugi ligands were less selective than the Triazine ligands for the hexapeptide. The triazine ligand 7,4 has been considered as the most selective for the designed affinity tag. In addition, preselected lead ligands for another hexapeptide (RW) of interest were studied. Mammalian cells HEK 293T were transfected with a vector expressing for GFP tagged with this peptide. The ligands immobilized onto agarose were loaded with cellular extracts, being noted that the lead A6C3 showed a high selectivity for the tag tested.

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16

Hoffman, Crystal Joyce. "Glucocorticoid Receptor Density and Binding Affinity in Horses with Systemic Inflammatory Response Syndrome". Thesis, Virginia Tech, 2014. http://hdl.handle.net/10919/48423.

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There were three objectives of this study. The first was to determine if commercially available fluorochromes could be used to determine the glucocorticoid receptor (GR) density and binding affinity (BA) in equine peripheral blood mononuclear cells. The second was to determine if there was a correlation between elevated plasma cortisol and GR density or binding affinity in healthy adult horses. The third objective was to evaluate the HPA axis in adult horses presenting with systemic inflammatory response syndrome (SIRS), and to determine where any alterations in HPA axis function occur in these patients compared to healthy adults. For the first part of the study, peripheral venous blood was collected from 3 healthy research horses on 3 days. Peripheral blood mononuclear cells were isolated using Ficoll gradient centrifugation. Phycoerythrin (PE)-CD44 was then used to extracellularly label leukocytes, and then an intracellular GR antibody was used to determine a baseline measurement of GR density and fluorescein isothiocyanate (FITC)-dexamethasone was used to determine binding affinity via flow cytometric analysis. Comparison of control samples to those for CD44, GR density, and GR binding affinity showed a statistically significant difference for all samples (P<0.0001, P<0.0001, and P<0.0001 respectively). This showed that the CD44, GR antibody, and FITC-dexamethasone could successfully be used to analyze equine peripheral blood mononuclear cells for GR activity. For the second part of the study, an ACTH stimulation test was performed on 8 healthy horses in order to induce an increase in endogenous cortisol production. Plasma cortisol levels, GR density, and GR binding affinity were measured at baseline, 4, 8, and 24 hours after treatment. Median basal cortisol concentration was 4.9, range 3.2-6.1 μg/dl. This initially increased following ACTH stimulation to 5.6, range 4.8-7.4 μg/dl, then showed a significant decrease by 8 hours post ACTH administration to 1.4, range 1.1-2.7 μg/dl (P=0.0221). No correlation was observed between plasma cortisol concentration in healthy horses and GR density or binding affinity (r=-0.145, P=0.428 and r=0.046, P=0.802, respectively). For the third phase of the study, horses (N=10) with systemic inflammatory response syndrome (SIRS) were compared to healthy, age and sex matched controls (N=10) presenting for lameness evaluation or ophthalmologic examination. Blood was collected from SIRS cases and controls on presentation to the Equine Medical Center. A CBC, serum biochemistry, and serum ACTH and cortisol measurements were performed. GR density and binding affinity were also determined. Nonsurvivors had a significantly decreased GR binding affinity (P=0.008) and demonstrated a trend towards an increase in the ACTH:cortisol ratio. ROC analysis was performed for serum ACTH and cortisol concentrations, the ACTH:cortisol ratio, GR density and GR binding affinity, and triglycerides to determine cut-off values associated with nonsurvival. These were then used to analyze this population using Fischer's exact test to determine the odds ratio (OR) associated with nonsurvival for each variable. This revealed that a serum triglyceride concentration greater than 28.5 mg/dl was associated with nonsurvival (OR=117, 95% CI, 1.94-7060). The other variables were not found to be significantly associated with nonsurvival, although a Delta BA% of less than 35.79% was found to be closely associated with nonsurvival (OR=30.33, 95% CI, 0.96-960.5). Additionally, a significant negative correlation was detected between the plasma ACTH concentration and Delta BA% (r=-0.685, P=0.029) and the ACTH:cortisol ratio and the Delta BA% (r=-0.697, P=0.025). This study showed that nonsurviving horses with SIRS had a significantly decreased GR binding affinity compared to survivors, and a tendency toward an increase in their ACTH:cortisol ratios. This confirms that HPA axis dysfunction occurs in adult horses with SIRS as tissue resistance to glucocorticoids, and potentially relative adrenal insufficiency as well. These results suggest that there are horses with SIRS that might benefit from "physiologic" doses of synthetic glucocorticoids to complement their relative adrenal insufficiency in addition to their poor tissue sensitivity. Further research should focus on methods to more rapidly determine which horses might benefit from treatment with glucocorticoids on presentation, as well as to more accurately determine prognosis for survival.
Master of Science

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17

Taylor, Eric S. "Molecular Investigations of the Epidermal Growth Factor Receptor and Its Affinity Toward Asbestos". The Ohio State University, 2012. http://rave.ohiolink.edu/etdc/view?acc_num=osu1324066851.

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18

Mehta, Ratnavali C. "Pharmacological evaluation of trimetoquinol analogs as affinity ligands for beta-adrenergic receptor systems /". The Ohio State University, 1997. http://rave.ohiolink.edu/etdc/view?acc_num=osu1487945015617968.

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19

林國泰 i Kwok-tai Lam. "Differential expression of p75 low affinity neurotrophin receptor in hypoxic-ischemic neonatal mouse brain". Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 1998. http://hub.hku.hk/bib/B29797822.

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20

Fear, David Jonathan. "Analysis of the high affinity IgE receptor (FcεRI) α-chain promoter, and its regulation". Thesis, King's College London (University of London), 1998. https://kclpure.kcl.ac.uk/portal/en/theses/analysis-of-the-high-affinity-ige-receptor-fceri-xchain-promoter-and-its-regulation(846c4e6a-5dad-40c4-aed5-4e5601944f66).html.

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21

Madian, Ashraf G. "Using affinity purification -- mass spectrometry to identify aryl hydrocarbon receptor nuclear translocator interacting proteins". Scholarly Commons, 2005. https://scholarlycommons.pacific.edu/uop_etds/605.

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The aryl hydrocarbon nuclear translocator (ARNT) belongs to the family of basic helix loop helix proteins. ARNT forms a heterodimer with aryl hydrocarbon receptor (AhR). This heterodimer binds to the dioxin responsive element (DRE) causing the regulation of the gene expression of some enzymes such as CYP1Al. Studies show that the ARNT-AhR heterodimer needs protein factors to bind to DRE, and most of these protein factors are still unknown. ARNT also heterodimerizes with hypoxia inducible factor 1 a (HIF-1 a) which mediates the cellular responses to hypoxia. The purpose of this research is to use a combination of affinity chromatography and mass spectrometry techniques to identify the proteins that interact with ARNT. We chose mouse liver as the protein source. We planned to isolate the ARNT interacting proteins from the mouse liver protein extract by using the TALON® resin column bounded with 6x HIS-ARNT. Two negative control experiments were done. The first one by the application of the liver extract only on non-ARNT bound column. The second one by the application of 100 mg of bovine serum albumin on 6x HIS-ARNT bound column. The mouse liver extract was applied on 6x HIS-ARNT bound column. The column was washed with an increasing concentration of potassium chloride (0.05 M- 1 M). ARNT was eluted with a buffer containing 250 and 500 mM imidazole. The different washing fractions were compared with the negative control experiments. There was no difference between this and experiment and negative controls. We also tested using in-vitro chemical cross-linking with formaldehyde. Some distorted bands that may '· represent crosslinked proteins appeared above ARNT molecular weight by the addition of 1% paraformaldehyde for 20 minutes at 37°C, and for 2 hours at 30° and room temperatures. These bands were absent in the negative control experiments. The mass spectrometric protocols for identification of trace amount of proteins using peptide mass fingerprinting were tested using a standard protein (Bovine Serum Albumin).

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22

Koide, Tomoaki. "Localization of trkB and low-affinity NGF receptor mRNA in the developing rat retina". Kyoto University, 1995. http://hdl.handle.net/2433/187158.

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本文データは平成22年度国立国会図書館の学位論文(博士)のデジタル化実施により作成された画像ファイルを基にpdf変換したものである.
Neuroscience Letters, vol.185(3), pp.483-186, 1995, doi:10.1016/0304-3940(95)11257-W.
Kyoto University (京都大学)
0048
新制・課程博士
博士(医学)
甲第5966号
医博第1632号
新制||医||602(附属図書館)
UT51-95-D285
京都大学大学院工学研究科脳統御医科学系専攻
(主査)教授井出千束, 教授本田孔士, 教授菊池晴彦
学位規則第4条第1項該当

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23

Lam, Kwok-tai. "Differential expression of p75 low affinity neurotrophin receptor in hypoxic-ischemic neonatal mouse brain /". Hong Kong : University of Hong Kong, 1998. http://sunzi.lib.hku.hk/hkuto/record.jsp?B21779156.

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24

Leib, Nicole [Verfasser]. "Functional cooperation of Toll-like receptor signaling and the high-affinity receptor for IgE, FcεRI, on human Langerhans cells / Nicole Leib". Bonn : Universitäts- und Landesbibliothek Bonn, 2018. http://d-nb.info/1153467194/34.

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25

Marsh, Andrew. "Characterisation of the type I IGF receptor binding surfaces of insulin-like growth factor 1 using protein engineering". Thesis, University of Newcastle Upon Tyne, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.245705.

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26

Bonnefoy, Jean-Yves. "The low affinity receptor for IgE on human B lymphocytes : detection, biochemical characterization and regulation". Lyon 1, 1987. http://www.theses.fr/1987LYO1T147.

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27

Hoshikawa, Hajime. "High affinity binding of oxidized LDL to mouse lectin-like oxidized LDL receptor (LOX-1)". Kyoto University, 1999. http://hdl.handle.net/2433/182282.

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28

Liang, Boying. "Studies of Ligand-Receptor Pairs Utilizing Polymerized Planar Supported Lipid Bilayers". Diss., The University of Arizona, 2013. http://hdl.handle.net/10150/311232.

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Artificial membranes composed of natural lipids are not stable when exposed to air/vacuum, surfactant, organic solvent, etc. Polymerizable lipids provide an opportunity to broaden the use of lipid membranes to study ligand-receptor pairs under harsh experimental conditions. This dissertation presents the utilization of polymerizable lipids in matrix assisted laser desorption and ionization-mass spectrometry (MALDI-TOF MS) for analysis of ligands bound to membrane receptors. This platform may be applied to rapid drug-screening for membrane receptors including transmembrane proteins. Bacterial toxins and their membrane receptors were used as model ligand-receptor pairs to demonstrate the feasibility of using polymerizable lipids to detect and identify ligands by MALDI-TOF MS. Cholera toxin B (CTB) was successfully detected bound to polymerized lipid membranes with incorporation of its membrane receptor, GM1, while no CTB was detected in non-polymerizable lipid membranes. This affinity capture platform based on poly(lipid) showed a high resistance to interferences. On-plate digestion of bound CTB was performed and 57% amino acid sequence coverage was achieved. Total internal reflection fluorescence microscopy (TIRF-M) was applied to compare CTB-GM1 binding affinity in polymerized and unpolymerized membranes. Under a static flow system, the binding between CTB and GM1 was found to be stronger in polymerized membranes than other membranes. However, the ligand concentration under a static flow system is not in excess and the apparent binding affinity is likely to be significantly different than the true value. The true binding affinity can be approached under a continuous flow system, however equilibration time was found to be too long to address experimentally. Membrane fluidity, which may be required to maintain the membrane receptor activity, is suppressed in poly(lipid) membranes compared to unpolymerized membranes. In order to maintain fluidity, a non-polymerizable lipid was mixed into a polymerized lipid. Fluorescence recovery after photobleaching (FRAP) data showed that fluidity of membrane composed of the mixed lipid was maintained compared to pure poly(lipid). Phase segregation of polymerized lipid and non-polymerizable lipid was detected by atomic force microscopy (AFM). CTB bound to GM1 in mixed lipid membranes was detected by MALDI-MS, indicating the mixed lipid membranes retain stability under MALDI-MS analysis conditions.

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29

Sharma, Kanika [Verfasser], i Sigrun [Gutachter] Korsching. "Olfactory coding in vertebrates: a novel tuning mechanism for receptor affinity and evolution of the olfactory receptor repertoire / Kanika Sharma ; Gutachter: Sigrun Korsching". Köln : Universitäts- und Stadtbibliothek Köln, 2018. http://d-nb.info/1204199361/34.

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30

Shu, Hong. "Design and Synthesis of CB1 Receptor Ligands and Synthesis of Amphibian Alkaloids". ScholarWorks@UNO, 2009. http://scholarworks.uno.edu/td/1104.

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Our project was aimed at the development of novel CB1 cannabinoid receptor antagonists that may have clinical applications for the treatment of cannabinoid and psychostimulant addiction. In this study, we designed, synthesized, and established the CB1 affinity for the 1,5-diaryl-1,2,3- triazole esters, a series of 4,5-diaryl-1-substituted-1,2,3-triazole analogues and a series of 4,5- diaryl-2-substituted-1,2,3-triazoles. Our research group has been interested in the synthesis of amphibian alkaloids due to their interesting biological activities. We have recently developed a general synthetic strategy which can rapidly prepare a few amphibian alkaloids simply from the abundant natural product (-)- cocaine This strategy was first successfully applied to the synthesis of (-)-monomorine. More recently, this strategy has also been utilized in the syntheses of both of the enantiomers of cispyrrolidine 225H and (+)-gephyrotoxin 287C.

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31

Redhu, Naresh Singh. "Molecular regulation and effector functions of the high affinity IgE receptor (FcεRI) in human airway smooth muscle cells". PLoS One (The Public Library of Science, USA), 2009. http://hdl.handle.net/1993/5305.

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The prevalence and economic burden of chronic airway disorders such as asthma is on the rise annually. Allergic asthma is characterized by chronic airway inflammation, airway hyper-responsiveness (AHR), and structural airway remodeling due to increased smooth muscle mass. Most allergic asthma occurs due to the overproduction of immunoglobulin E (IgE) antibodies against common allergens. Classically, IgE has been shown to modulate airway smooth muscle (ASM) contraction/relaxation which is believed to be the underlying cause of airway hyperreactivity. However, the molecular mechanisms underlying IgE effects on ASM cell are not established. Recently, the high-affinity Fc receptor for IgE (FcεRI) has been identified in human ASM cells in vitro and in vivo within bronchial biopsies of allergic asthma patients. However, it is unknown whether FcεRI activation on ASM can modulate the immune response within the airways. We hypothesized that the IgE-FcεRI interaction plays a key role in inducing phenotypic and functional changes in ASM cells that eventually contributes to the establishment of airway inflammation, AHR, and remodeling. We sought to investigate the regulation, effector functions, and underlying mechanisms of FcεRI activation in ASM cells. Our work shows that the proinflammatory tumor necrosis factor (TNF) and T helper type 2 (Th2) cytokine interleukin (IL)-4 enhanced the FcεRI abundance and amplified the IgE-induced chemokine (eotaxin-1/CCL11, RANTES/CCL5, IL-8/CXCL8, and IP-10/CXCL10) release in ASM cells via transcriptional mechanisms. Both TNF and IgE induced a novel, Th2-favoring cytokine thymic stromal lymphopoietin (TSLP) through the activation of spleen tyrosine kinase (Syk), and nuclear factor kappa B (NF-κB) and activator protein-1 (AP-1). In addition, IgE induced de novo DNA synthesis and ASM cell proliferation via mitogen-activated protein kinases (MAPKs) and signal-transducer and activator of transcription 3 (STAT3) activation. Collectively, our data suggest that the IgE-induced FcεRI activation leads to the expression of multiple chemokines in ASM which may indirectly recruit inflammatory cells and promote allergic airway inflammation; IgE induces TSLP which can promote the Th2 immune responses within the airways; and IgE may potentially induce airway remodeling by directly inducing ASM cell proliferation. Therefore, targeting the IgE-FcεRI network on ASM may offer a novel therapeutic strategy in allergic asthma.

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32

Khan, Samina E. "An investigation of structure activity effects of D-ring substitution of estradiol on estrogen receptor affinity". Thesis, Loughborough University, 1992. https://dspace.lboro.ac.uk/2134/28192.

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The 16α-alkylation was achieved via treatment of 3-methoxyestra-1,3,5(10)-trien-17-one-N,N-dimethylhydrazone (118) with n-butyllithium and the appropriate haloalkane to afford exclusive 16α-substitution (119). Subsequent, cupric ion-catalysed hydrolysis, 17-ketone reduction and removal of the 3-hydroxyl protecting group, furnished 16α-methylestra-1,3,5(10)-trien-3,17β-diol (122a) and 16α-ethylestra-1,3,5(10)-trien-3,17β-diol (122b). An alternative sequence to the 16α-ligands, by direct alkylation of the 17-keto-estrone enolate, was also investigated. In this manner 16α-allylestra-1,3,5(10)-trien-3,17β-diol (122c) and 16α-benzylestra-1,3,5,(10)-trien-3,17β-diol (122d) were obtained.

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33

Savdie, Cheryl. "Characterization of the cellular and molecular determinants for the internalization of the high affinity neurotensin receptor". Thesis, McGill University, 2002. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=29473.

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Neurotensin is a thirteen amino acid peptide that is found both in the central and peripheral nervous systems. In the CNS, it controls the synthesis and release of dopamine, modulates anterior pituitary hormone secretion, and provides non-opioid-receptor-dependent analgesia. In the gut, it modulates gastric acid secretion, pancreatic secretion, and colonic motility. In the immune system, it modulates IL-1 and mast cell secretion and stimulates macrophage phagocytosis. Neurotensin also acts as a growth factor for normal and cancer cells. Neurotensin mediates its effects through three neurotensin receptors: NTR1, NTR2, and NTR3. NTR1 and NTR2 belong to the G-protein coupled receptor (GPCR) family, while NTR3 is a single transmembrane receptor identical to sortilin. NTR1 has the highest affinity for neurotensin (Kd of 0.1--0.3 nM). Internalization of GPCRs, including NTR1 and NTR2, was shown to play an important role in initiating signaling activity as well as in desensitizing and resensitizing the receptors.
In this study, the internalization of the NTR1 was investigated. (Abstract shortened by UMI.)

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34

Linhult, Martin. "Protein engineering to explore and improve affinity ligands". Doctoral thesis, KTH, Biotechnology, 2003. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-3632.

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In order to produce predictable and robust systems forprotein purification and detection, well characterized, small,folded domains descending from bacterial receptors have beenused. These bacterial receptors, staphylococcal protein A (SPA)and streptococcal protein G (SPG), possess high affinity to IgGand / or HSA. They are composed of repetitive units in whicheach one binds the ligand independently. The domains foldindependently and are very stable. Since the domains also havewellknown three-dimensional structures and do not containcysteine residues, they are very suitable as frameworks forfurther protein engineering.

Streptococcal protein G (SPG) is a multidomain proteinpresent on the cell surface ofStreptococcus. X-ray crystallography has been used todetermine the binding site of the Ig-binding domain. In thisthesis the region responsible for the HSA affinity of ABD3 hasbeen determined by directed mutagenesis followed by functionaland structural analysis. The analysis shows that the HSAbindinginvolves residues mainly in the second α-helix.

Most protein-based affinity chromatography media are verysensitive towards alkaline treatment, which is the preferredmethod for regeneration and removal of contaminants from thepurification devices in industrial applications. Here, aprotein engineering strategy has been used to improve thetolerance to alkaline conditions of different domains fromprotein G, ABD3 and C2. Amino acids known to be susceptibletowards high pH were substituted for less alkali susceptibleresidues. The new, engineered variants of C2 and ABD shownhigher stability towards alkaline pH. Also, very important forthe potential use as affinity ligands, these mutated variantsretained the secondary structure and the affinity to HSA andIgG, respectively. Moreover, dimerization was performed toinvestigate whether a higher binding capacity could be obtainedby multivalency. For ABD, binding studies showed that divalentligands coupled using non-directed chemistry demonstrated anincreased molar binding capacity compared to monovalentligands. In contrast, equal molar binding capacities wereobserved for both types of ligands when using a directed ligandcoupling chemistry involving the introduction and recruitmentof a unique C-terminal cysteine residue.

The staphylococcal protein A-derived domain Z is also a wellknown and thoroughly characterized fusion partner widely usedin affinity chromatography systems. This domain is consideredto be relatively tolerant towards alkaline conditions.Nevertheless, it is desirable to further improve the stabilityin order to enable an SPA-based affinity medium to withstandeven longer exposure to the harsh conditions associated withcleaning in place (CIP) procedures. For this purpose adifferent protein engineering strategy was employed. Smallchanges in stability due to the mutations would be difficult toassess. Hence, in order to enable detection of improvementsregarding the alkaline resistance of the Z domain, a by-passmutagenesis strategy was utilized, where a mutated structurallydestabilized variant, Z(F30A) was used as a surrogateframework. All eight asparagines in the domain were exchangedone-by-one. The residues were all shown to have differentimpact on the alkaline tolerance of the domain. By exchangingasparagine 23 for a threonine we were able to remarkablyincrease the stability of the Z(F30A)-domain towards alkalineconditions. Also, when grafting the N23T mutation to the Zscaffold we were able to detect an increased tolerance towardsalkaline treatment compared to the native Z molecule. In allcases, the most sensitive asparagines were found to be locatedin the loops region.

In summary, the work presented in this thesis shows theusefulness of protein engineering strategies, both to explorethe importance of different amino acids regarding stability andfunctionality and to improve the characteristics of aprotein.

Keywords:binding, affinity, human serum albumin (HSA),albumin-binding domain (ABD), affinity chromatography,deamidation, protein A, stabilization, Z-domain, capacity,protein G, cleaning-in-place (CIP), protein engineering, C2receptor.

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35

Sato, Sumito. "High-affinity urokinase-derived cyclic peptides inhibiting urokinase-urokinase receptor-interaction : effects on tumor growth and spread". kostenfrei, 2009. https://mediatum2.ub.tum.de/node?id=653320.

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36

Shenderov, Eugene. "In vivo and In vitro Studies of T-Cell Receptor-ligland-MHC Affinity and T-Cell Function". Thesis, University of Oxford, 2009. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.504593.

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37

Kramer, A. M. "Delineating the impact of binding-domain affinity and kinetic properties on Chimeric Antigen Receptor T-cell function". Thesis, University College London (University of London), 2017. http://discovery.ucl.ac.uk/1558284/.

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CD19 Chimeric Antigen Receptor (CAR) therapy represents a breakthrough in the treatment of relapsed/refractory acute lymphoblastic leukaemia and early-phase clinical trials with CAR-modified-T cells have shown unprecedented responses. A CAR is a recombinant receptor that combines a single chain variable fragment (scFv) against a tumour-associated antigen and an intracellular activation domain of the T cell receptor (TCR), recognizing membrane-bound antigen, typically with a higher affinity compared to TCR-pMHC affinity. We compared the influences of differences in on- and off-rates underlying the binding kinetics of CD19 CARs on downstream CAR T cell responses and constructed a novel CAR derived from the CAT hybridoma demonstrating a lower affinity as a result of a greater off-rate, whilst maintaining an on-rate nearly identical to FMC63, the scFv most-used in clinical trials. Using in vitro pre-clinical models we demonstrated that CAT-CAR+ T cells showed increased proliferation, higher cytotoxic responses, and increased cytokine production, as well as a greater number of interactions between CAT-CAR+ T cells and target cells and a greater motility in comparison to FMC63. In vivo CAT-CAR+ T cells showed an enhanced ability to clear disease, proliferate and produce cytokines, displaying markers characteristic for improved T cell fitness. We believe that, analogous to the natural TCR, a lower overall binding affinity might be mitigated by a relatively faster off-rate in the setting of a constant on-rate and propose that this may enhance CAR T cell function through serial triggering, where increased target:effector interaction time, may lead to exhaustion and activation induced cell death. These data have important implications for the development of future CARs.

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38

Fournet, Steven P. "High Resolution X-ray Diffraction Analysis of CB1 Receptor Antagonists as a Means to Explore Binding Affinity". ScholarWorks@UNO, 2013. http://scholarworks.uno.edu/td/1737.

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Abstract Charge density studies have been conducted on ten CB1 cannabinoid receptor antagonists via high resolution x-ray crystallography. Bond critical point values and various other properties derived from these studies including the electrostatic potential were analyzed in correlation to the affinity of each compound with the CB1 receptor. Correlation/anti-correlation was found between several properties and Ki. The data was also interpreted by principal component analysis with three principal components accounting for 85% of the data variation. Data mining was limit due to the low sample count and the requirements set for the inclusion of correlated/anti-correlated variables left fewer variables to analyze. The model presented is left for future interpretation.

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39

Paul, Alan Burnett. "β-NGF and its low-affinity receptor (p75LNGFR) in benign prostatic hyperplasia and adenocarcinoma of the prostate". Thesis, University of Edinburgh, 1999. http://hdl.handle.net/1842/29936.

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β-NGF was measured in human benign prostatic hyperplastic (BPH), adenocarcinomatous and normal tissues using an enzyme-linked immunosorbent assay. Concentrations were 1992pg/g wet weight in BPH tissue (SD = 684pg/g), 3100pg/g in adenocarcinomatous tissue (SD = 1503pg/g), and 2690pg/g in normal tissue. mRNA transcripts for β-NGF were demonstrated in prostate tissues by reverse transcription-polymerase chain reaction showing that the β-NGF measured in prostate tissue was endogenously produced. Western blotting allowed the demonstration that immunoreactive β-NGF in the prostate represented dimeric β-NGF and not behaviour β-NGF-like proteins which have been described elsewhere. Immunohistochemistry for β-NGF localised the hormone to the prostate epithelium in BPH, cancer and normal tissue. This epithelial localisation was confirmed by video-assisted tissue morphometric studies. Thereby it was shown that specimen β-NGF concentrations correlated well and statistically significantly with specimen prostatic glandular content. Morphometric studies also allowed the expression of β-NGF concentrations in terms of each specimen's contained, β-NGF secreting, glandular tissue. Thereby it was demonstrated that BPH glandular tissue produced greater concentrations of β-NGF (1597pg/100mg glandular tissue, SD = 788pg/100mg) than did malignant glandular tissue (1058pg/100mg glandular tissue, SD = 559pg/100mg), in spite of the higher concentrations of β-NGF found grossly in adenocarcinomatous tissue. β-NGF concentrations did not correlate with differential of adenocarcinomas or with the degree of neurodocrine differentiation therein. The thesis suggests that human prostatic β-NGF has a role similar to that described in other tissues. That is the maintenance and stimulation of adrenergic nerves. This may be relevant to the widespread use of sympatholytic drugs in the treatment of BPH.

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40

Mockford, Esther Helen. "The identification and expression of a novel variant of the beta subunit of the high affinity IgE receptor". Thesis, University of Oxford, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.312622.

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41

McCarthy-Troke, Melanie. "Comparative modelling and mutational analysis of the alpha-subunit of the high affinity receptor for IgG, FcÎ³RI". Thesis, University of Glasgow, 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.399675.

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42

Zloh, Mire. "Conformational studies of the domains and subunits of the high affinity IgE receptor by NMR and molecular modelling". Thesis, University College London (University of London), 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.312946.

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43

Pardoe, David Alan. "Synthetic chemistry and protein molecular modelling towards novel, high affinity ligands for the 5-HTâ†1â†E receptor". Thesis, University of Bristol, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.388103.

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44

Friedman, Mikaela. "Affibody molecules targeting the epidermal growth factor receptor for tumor imaging applications". Doctoral thesis, Stockholm : School of Biotechnology, Royal Institute of Technology (KTH), 2008. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-4710.

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45

Krauchunas, Amber R. M. "CD8⁺ T-cell response potential, as determined by expression of the high affinity interleukin-2 receptor, in murine AIDS /". Connect to online version, 2006. http://ada.mtholyoke.edu/setr/websrc/pdfs/www/2006/149.pdf.

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46

Ferri, Catharine C. "The role of the p75 low-affinity neurotrophin receptor in the peripheral and central nervous systems following nerve injury". Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1998. http://www.collectionscanada.ca/obj/s4/f2/dsk2/ftp01/MQ31203.pdf.

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47

Sihra, Bhupinder Singh. "Studies investigating peripheral blood derived cells that express the high affinity receptor for immunoglobulin E (FceRI) in allergic disorders". Thesis, University of Southampton, 2008. https://eprints.soton.ac.uk/67619/.

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It is just forty years since the identification of immunoglobulin E (IgE) as the reagin responsible for allergen induced immediate hypersensitivity reactions. IgE exerts its biological actions through the binding of its Fc fragment to specific Fc receptors on effector cells. There are two predominant Fc receptors for IgE – FcεRI, which has a very high affinity for IgE and FcεRII, which shows less avid binding. For much of the first two decades after the identification of IgE, it was thought that FcεRI expression was limited to mast cells and basophils and that IgE binding to other cell types such as Blymphocytes and antigen presenting cells (APCs) was mainly due to FcεRII. However with major advances in characterisation and functional knowledge of FcεRI, particularly in the last fifteen years, it has become apparent that FcεRI can be expressed on several more cell types that may be involved in initiation and maintenance of allergic inflammation – including APCs (monocytes and dendritic cells) and possibly eosinophils. The research described in the four papers forming this thesis was completed during this period and evaluated FcεRI expression on different cell types, their potential roles in allergen induced inflammatory responses and whether successful therapeutic strategies for allergic disorders may involve actions on FcεRI+ cells. The relative expression of FcεRI on peripheral blood basophils, monocytes and eosinophils from atopic and non-atopic subjects and any relationship with serum IgE concentrations was assessed in the first paper. The second study examined a potentially important role for basophils as a cellular source of rapidly releasable IL-4 which may help initiate allergen induced TH2 responses. The next study investigated the possible effects on allergen induced early and late asthmatic responses of the immunosuppressive drug cyclosporin A which had been shown both to inhibit mast cell and basophil degranulation and cytokine secretion (particularly by CD4+ T-cells). The final study evaluated FcεRI expression on these cell types as well humoral factors (e.g. seasonal changes in allergen specific IgG and IgE) in subjects who, after 3 to 4 years of grass pollen immunotherapy, had continued on either active or placebo immunotherapy for a further 3 years. A historical perspective explaining some of the reasons the studies were done is provided in the introductory chapter whilst the discussion chapter at the end reviews how many of the findings of the study have evolved in subsequent years right up to the present day and finishes off with a brief synopsis of how rapidly increasing knowledge of the regulatory functions of dendritic cells (expressing FcεRI and often monocyte derived) has resulted in better understanding of the mechanisms of allergen specific immunotherapy and is leading to more effective treatment modalities.

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48

Sarpong, Yaw Acheampong. "Influence of HMGB1 on Nucleosome Structure and Estrogen Receptor Binding Affinity to Concensus Estrogen Response Element on Nucleosomal DNA". Bowling Green State University / OhioLINK, 2010. http://rave.ohiolink.edu/etdc/view?acc_num=bgsu1288629979.

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SARPONG, YAW A. "THE BINDING OF ESTROGEN, PROGESTERONE AND GLUCOCORTICOID RECEPTORS TO THEIR RECOGNITION SITES IN A NUCLEOSOME AND THE EFFECT OF HMGB1 ON THE BINDING AFFINITY". Bowling Green State University / OhioLINK, 2006. http://rave.ohiolink.edu/etdc/view?acc_num=bgsu1154368368.

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50

Watson, Diana Ruth. "The role of ASP 195 in the transmembrane domain of the #alpha# subunit of the high affinity receptor for IgE". Thesis, University of Sheffield, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.312290.

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