Record Details

Biophysical studies of ligand-gated ion channels [electronic resource] / Betty Ko Wong ; Dennis A. Dougherty, advisor ; Henry A. Lester, co-advisor.
Pasadena, California : California Institute of Technology, 2017.
1 online resource (xi, 110 leaves) : digital (13 Mb), illustrations (some color).
CIT theses ; 2017
This dissertation describes building a methodology for and the biophysical studies of ligand-gated ion channels (LGICs). The primary focus of the first half of this dissertation is on developing a fluorescence-based assay to broadly study LGICs. Chapter 2 describes the site-selective incorporation of a turn-on fluorophore via unnatural amino acid mutagenesis on the mouse muscle-type nicotinic acetylcholine receptor (nAChR) in Xenopus oocytes as a proof-of-principle study. This method has proven to yield very low levels of undesired fluorescent background, which was a problem for previous incorporation techniques. Chapter 3 describes efforts towards imaging this in vivo system using lifetime imaging with efforts hampered by the inability to detect a clear signal. Chapter 4 describes efforts to apply the lifetime imaging approach towards a different system involving 5-HT3 proteins fused to fluorescent proteins in COS-7 cells. The second half of this dissertation focuses on studies of menthol, a flavorant added to cigarettes that contributes to smoking addiction, as a negative allosteric modulator of the ł ø42 nAChR. Chapter 5 reveals the stereochemical effects, or rather lack of, of menthol on the two stoichiometries of the ł ø42 receptor. Chapter 6 seeks to identify the residue interactions with menthol of the ł ø42 receptor using a combination of computational and experimental studies.
Advisor and committee chair names found in the thesis' metadata record in the digital repository.
Dissertation note:
Thesis (Ph. D.) -- California Institute of Technology, 2017.
Bibliography, etc. note:
Includes bibliographical references.
Linked resources:
Caltech Connect
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 Record created 2017-11-08, last modified 2017-11-08

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