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Binding Studies of Neuronal Nicotinic Acetylcholine Receptors Expressing Unnatural Amino Acids

Citation

Da Silva Tavares, Ximena (2015) Binding Studies of Neuronal Nicotinic Acetylcholine Receptors Expressing Unnatural Amino Acids. Dissertation (Ph.D.), California Institute of Technology. doi:10.7907/Z9NC5Z45. http://resolver.caltech.edu/CaltechTHESIS:04242015-121623394

Abstract

Nicotinic acetylcholine receptors are pentameric ligand-gated ion channels mediating fast synaptic transmission throughout the peripheral and central nervous systems. They have been implicated in various processes related to cognitive functions, learning and memory, arousal, reward, motor control and analgesia. Therefore, these receptors present alluring potential therapeutic targets for the treatment of pain, epilepsy, Alzheimer’s disease, Parkinson’s disease, Tourette’s syndrome, schizophrenia, anxiety, depression and nicotine addiction. The work detailed in this thesis focuses on binding studies of neuronal nicotinic receptors and aims to further our knowledge of subtype specific functional and structural information.

Chapter 1 is an introductory chapter describing the structure and function of nicotinic acetylcholine receptors as well as the methodologies used for the dissertation work described herein. There are several different subtypes of nicotinic acetylcholine receptors known to date and the subtle variations in their structure and function present a challenging area of study. The work presented in this thesis deals specifically with the α4β2 subtype of nicotinic acetylcholine receptor. This subtype assembles into 2 closely related stoichiometries, termed throughout this thesis as A3B2 and A2B3 after their respective subunit composition. Chapter 2 describes binding studies of select nicotinic agonists on A3B2 and A2B3 receptors determined by whole-cell recording. Three key binding interactions, a cation-π and two hydrogen bonds, were probed for four nicotinic agonists, acetylcholine, nicotine, smoking cessation drug varenicline (Chantix®) and the related natural product cytisine.

Results from the binding studies presented in Chapter 2 show that the major difference in binding of these four agonists to A3B2 and A2B3 receptors lies in one of the two hydrogen bond interactions where the agonist acts as the hydrogen bond acceptor and the backbone NH of a conserved leucine residue in the receptor acts as the hydrogen bond donor. Chapter 3 focuses on studying the effect of modulating the hydrogen bond acceptor ability of nicotine and epibatidine on A3B2 receptor function determined by whole-cell recording. Finally, Chapter 4 describes single-channel recording studies of varenicline binding to A2B3 and A3B2 receptors.

Item Type:Thesis (Dissertation (Ph.D.))
Subject Keywords:nicotinic acetylcholine receptors, unnatural amino acids, nicotine, varenicline, single-channel
Degree Grantor:California Institute of Technology
Division:Chemistry and Chemical Engineering
Major Option:Chemistry
Thesis Availability:Public (worldwide access)
Research Advisor(s):
  • Dougherty, Dennis A.
Thesis Committee:
  • Reisman, Sarah E. (chair)
  • Dervan, Peter B.
  • Tirrell, David A.
  • Dougherty, Dennis A.
Defense Date:5 August 2014
Non-Caltech Author Email:ximedasilva (AT) gmail.com
Funders:
Funding AgencyGrant Number
NIHNS 34407
NIHNS 11756
California Tobacco-Related Disease Research Program16RT-0160
Record Number:CaltechTHESIS:04242015-121623394
Persistent URL:http://resolver.caltech.edu/CaltechTHESIS:04242015-121623394
DOI:10.7907/Z9NC5Z45
Default Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:8832
Collection:CaltechTHESIS
Deposited By: Ximena Da Silva Tavares Bongoll
Deposited On:01 May 2015 17:28
Last Modified:03 Feb 2016 23:20

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