CaltechAUTHORS
  A Caltech Library Service

Using Mutant Cycle Analysis to Elucidate Long-Range Functional Coupling in Allosteric Receptors

Shanata, Jai A. P. and Frazier, Shawnalea J. and Lester, Henry A. and Dougherty, Dennis A. (2011) Using Mutant Cycle Analysis to Elucidate Long-Range Functional Coupling in Allosteric Receptors. In: Allostery: Methods and Protocols. Methods in Molecular Biology. No.796. Springer , pp. 97-113. ISBN 9781617793332. PMCID PMC4006985. https://resolver.caltech.edu/CaltechAUTHORS:20111114-081228635

[img] PDF - Accepted Version
See Usage Policy.

1MB

Use this Persistent URL to link to this item: https://resolver.caltech.edu/CaltechAUTHORS:20111114-081228635

Abstract

Functional coupling of residues that are far apart in space is the quintessential property of allosteric receptors. Data from functional studies of allosteric receptors, such as whole-cell dose–response relations, can be used to determine if mutation to a receptor significantly impacts agonist potency. However, the classification of perturbations as primarily impacting binding or allosteric function is more challenging, often requiring detailed kinetic studies. This protocol describes a simple strategy, derived from mutant cycle analysis, for elucidating long-range functional coupling in allosteric receptors (ELFCAR). Introduction of a gain-of-function reporter mutation, followed by a mutant cycle analysis of the readily measured macroscopic EC_(50) values can provide insight into the role of many physically distant targets. This new method should find broad application in determining the functional roles of residues in allosteric receptors.


Item Type:Book Section
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1007/978-1-61779-334-9_6DOIArticle
http://www.springerlink.com/content/n6u6300h25487215/PublisherArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4006985PubMed CentralArticle
ORCID:
AuthorORCID
Lester, Henry A.0000-0002-5470-5255
Dougherty, Dennis A.0000-0003-1464-2461
Additional Information:© 2012 Springer Science+Business Media, LLC. We thank Kristin Rule Gleitsman for essential discussions throughout the development of ELFCAR. This work was supported by the National Institutes of Health (NS 34407; NS 11756). J.A.P.S. and S.F. were partially supported by National Research Service Awards from the NIH.
Funders:
Funding AgencyGrant Number
NIHNS 34407
NIHNS 11756
Subject Keywords:Allostery, Signal transduction, Conformational change, Coupling, Structure–function study, Ion channel, Nicotinic receptor, Amino acids
Series Name:Methods in Molecular Biology
Issue or Number:796
PubMed Central ID:PMC4006985
DOI:10.1007/978-1-61779-334-9_6
Record Number:CaltechAUTHORS:20111114-081228635
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20111114-081228635
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:27764
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:28 Nov 2011 22:01
Last Modified:09 Nov 2021 16:51

Repository Staff Only: item control page