Publication Date
2012
Document Type
Thesis
Committee Members
Thomas Brown (Committee Member), Adrian Corbett (Committee Member), J. Ashot Kozak (Advisor), Robert Putnam (Committee Member)
Degree Name
Master of Science (MS)
Abstract
The Orai3 protein forms a Ca2+ channel with a pharmacological profile distinct from its close relative, the store-operated Ca2+ channel Orai1. Though closely related to Orai1, the function of Orai3 in humans is still unclear. This study attempts to contribute to the body of knowledge by undertaking a pharmacological analysis of Orai3 in insect cells. We describe here the creation of a vector capable of expressing the mammalian Orai3 gene on an insect background. We demonstrate the ability to induce gene expression of Orai3 in these insect cells, and then assess the effectiveness of this system by characterizing the pharmacological properties with the drug 2-aminoethoxydiphenyl borate (2-APB). The results show that the current strategy used to express Orai3 will require additional refinement before the system can be considered generally useful for pharmacological studies, because expression of Orai3 may be affected by native proteins. The interference of expression seems confined to Orai Ca2+ channels, as mammalian STIM1 was also expressed and a response to 2-APB, albeit unexpected, was observed.
Page Count
66
Department or Program
Department of Neuroscience, Cell Biology & Physiology
Year Degree Awarded
2012
Copyright
Copyright 2012, some rights reserved. My ETD may be copied and distributed only for non-commercial purposes and may not be modified. All use must give me credit as the original author.
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.
