Effects of B2-Subunit Cytoplasmic-Loop Mutations A-4 Subunit Gain of Function Mutations on CNS Nicotinic Acetylcholine Receptors

Persistent Link:
http://hdl.handle.net/10150/297650
Title:
Effects of B2-Subunit Cytoplasmic-Loop Mutations A-4 Subunit Gain of Function Mutations on CNS Nicotinic Acetylcholine Receptors
Author:
Hossain, Farhana
Issue Date:
2013
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:
The effects of the β2-365AAQA368 cytoplasmic-loop mutant subunit and the α4L9’s gain-offunction mutant subunit on CNS Nicotinic Acetylcholine Receptors were determined in this experiment. Four HSP β2-365AAQA368 mutants in pCEP4 were transfected into human SH-EPI cells. These constructs were screened for function through both an 3H-Epibatidine in situ binding assay and a 86-Rb+ efflux assay. The gain of function mutant α4L9’s position 5 (β3β2Aα6/3β2α4L9’s-pSHE) was injected into Xenopus laevis oocytes and tested for function via a Two-Electrode-Voltage Clamp apparatus. The β2-365AAQA368 mutant caused a rightward shift of the EC50 relative to the wild-type. Three monoclones from the β2- 365AAQA368 mutant 86-Rb+ efflux assay showed at least some activity: clones 212, 117, and 213. The maximum function achieved was 1023 scpm (0.9%) in clone 213, but this was not enough function for drug discovery work. The gain of function mutant α4L9’s position 5 caused a leftward shift of the EC50 relative to the wild-type. The mutation also increased the amount of function of the receptor by at least 1.5 fold.
Type:
text; Electronic Thesis
Degree Name:
B.S.
Degree Level:
bachelors
Degree Program:
Honors College; Biology
Degree Grantor:
University of Arizona
Advisor:
Lukas, Ronald

Full metadata record

DC FieldValue Language
dc.language.isoenen_US
dc.titleEffects of B2-Subunit Cytoplasmic-Loop Mutations A-4 Subunit Gain of Function Mutations on CNS Nicotinic Acetylcholine Receptorsen_US
dc.creatorHossain, Farhanaen_US
dc.contributor.authorHossain, Farhanaen_US
dc.date.issued2013-
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.abstractThe effects of the β2-365AAQA368 cytoplasmic-loop mutant subunit and the α4L9’s gain-offunction mutant subunit on CNS Nicotinic Acetylcholine Receptors were determined in this experiment. Four HSP β2-365AAQA368 mutants in pCEP4 were transfected into human SH-EPI cells. These constructs were screened for function through both an 3H-Epibatidine in situ binding assay and a 86-Rb+ efflux assay. The gain of function mutant α4L9’s position 5 (β3β2Aα6/3β2α4L9’s-pSHE) was injected into Xenopus laevis oocytes and tested for function via a Two-Electrode-Voltage Clamp apparatus. The β2-365AAQA368 mutant caused a rightward shift of the EC50 relative to the wild-type. Three monoclones from the β2- 365AAQA368 mutant 86-Rb+ efflux assay showed at least some activity: clones 212, 117, and 213. The maximum function achieved was 1023 scpm (0.9%) in clone 213, but this was not enough function for drug discovery work. The gain of function mutant α4L9’s position 5 caused a leftward shift of the EC50 relative to the wild-type. The mutation also increased the amount of function of the receptor by at least 1.5 fold.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.disciplineBiologyen_US
thesis.degree.grantorUniversity of Arizonaen_US
dc.contributor.advisorLukas, Ronald-
All Items in UA Campus Repository are protected by copyright, with all rights reserved, unless otherwise indicated.