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dc.contributor.advisorLukas, Ronald
dc.contributor.authorHossain, Farhana*
dc.creatorHossain, Farhanaen_US
dc.date.accessioned2013-08-08T18:12:38Z
dc.date.available2013-08-08T18:12:38Z
dc.date.issued2013
dc.identifier.urihttp://hdl.handle.net/10150/297650
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.
dc.language.isoenen_US
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.titleEffects of B2-Subunit Cytoplasmic-Loop Mutations A-4 Subunit Gain of Function Mutations on CNS Nicotinic Acetylcholine Receptorsen_US
dc.typetexten_US
dc.typeElectronic Thesisen_US
thesis.degree.grantorUniversity of Arizonaen_US
thesis.degree.levelbachelorsen_US
thesis.degree.disciplineHonors Collegeen_US
thesis.degree.disciplineBiologyen_US
thesis.degree.nameB.S.en_US
refterms.dateFOA2018-08-30T09:47:29Z
html.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.


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