Binding Studies of Membrane Receptor CD47 with the C-terminal Domain of Thrombospondin-1

Persistent Link:
http://hdl.handle.net/10150/146078
Title:
Binding Studies of Membrane Receptor CD47 with the C-terminal Domain of Thrombospondin-1
Author:
Yang, Kimberly M.
Issue Date:
May-2010
Publisher:
The University of Arizona.
Rights:
Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
Abstract:
Secreted glycoprotein Thrombospondin-1 (TSP1), among its many functions, binds to membrane receptor CD47 in order to downregulate the nitric oxide pathway via inhibition of soluble guanylate cyclase (sGC). Physiologically, this process is anti-angiogenic and also affects vasodilation, platelet aggregation and tissue ischemia. Aside from the key players TSP1, CD47 and sGC, little is known about the mechanism of this pathway. This project seeks to understand the binding interaction between trimeric TSP1 and transmembrane receptor CD47. It is hypothesized that the C-terminal binding domain of TSP1 (E3CaG1) and the N-terminal extracellular domain of CD47 (CD47-NT) are sufficient to mediate binding. E3CaG1 was expressed and purified from Sf9 insect cells while CD47-NT was expressed and refolded from inclusion bodies in E. coli. Binding was tested via co-immunoprecipitation experiments utilizing both nickel and GST affinity resins. It was found that the GST- and His-tagged constructs of CD47-NT did not show any interaction with E3CaG1. While E3CaG1 has been shown to be fully active, further work is underway to determine whether CD47-NT is also properly folded and active.
Type:
text; Electronic Thesis
Degree Name:
B.S.
Degree Level:
bachelors
Degree Program:
Honors College; Biochemistry and Molecular Biophysics
Degree Grantor:
University of Arizona

Full metadata record

DC FieldValue Language
dc.language.isoenen_US
dc.titleBinding Studies of Membrane Receptor CD47 with the C-terminal Domain of Thrombospondin-1en_US
dc.creatorYang, Kimberly M.en_US
dc.contributor.authorYang, Kimberly M.en_US
dc.date.issued2010-05-
dc.publisherThe University of Arizona.en_US
dc.rightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.en_US
dc.description.abstractSecreted glycoprotein Thrombospondin-1 (TSP1), among its many functions, binds to membrane receptor CD47 in order to downregulate the nitric oxide pathway via inhibition of soluble guanylate cyclase (sGC). Physiologically, this process is anti-angiogenic and also affects vasodilation, platelet aggregation and tissue ischemia. Aside from the key players TSP1, CD47 and sGC, little is known about the mechanism of this pathway. This project seeks to understand the binding interaction between trimeric TSP1 and transmembrane receptor CD47. It is hypothesized that the C-terminal binding domain of TSP1 (E3CaG1) and the N-terminal extracellular domain of CD47 (CD47-NT) are sufficient to mediate binding. E3CaG1 was expressed and purified from Sf9 insect cells while CD47-NT was expressed and refolded from inclusion bodies in E. coli. Binding was tested via co-immunoprecipitation experiments utilizing both nickel and GST affinity resins. It was found that the GST- and His-tagged constructs of CD47-NT did not show any interaction with E3CaG1. While E3CaG1 has been shown to be fully active, further work is underway to determine whether CD47-NT is also properly folded and active.en_US
dc.typetexten_US
dc.typeElectronic Thesisen_US
thesis.degree.nameB.S.en_US
thesis.degree.levelbachelorsen_US
thesis.degree.disciplineHonors Collegeen_US
thesis.degree.disciplineBiochemistry and Molecular Biophysicsen_US
thesis.degree.grantorUniversity of Arizonaen_US
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