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PMID: 20637420 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Conformational changes during the gating of a potassium channel revealed by structural mass spectrometry.

Structure (London, England : 1993) ·Vol. 18 ·No. 7 ·2010-07-14 ·Pages 839-46

Gupta S, Bavro VN, D'Mello R, Tucker SJ, Vénien-Bryan C, Chance MR

Abstract

Potassium channels are dynamic proteins that undergo large conformational changes to regulate the flow of K(+) ions across the cell membrane. Understanding the gating mechanism of these channels therefore requires methods for probing channel structure in both their open and closed conformations. Radiolytic footprinting is used to study the gating mechanism of the inwardly-rectifying potassium channel KirBac3.1. The purified protein stabilized in either open or closed conformations was exposed to focused synchrotron X-ray beams on millisecond timescales to modify solvent accessible amino acid side chains. These modifications were identified and quantified using high-resolution mass spectrometry. The differences observed between the closed and open states were then used to reveal local conformational changes that occur during channel gating. The results provide support for a proposed gating mechanism of the Kir channel and demonstrate a method of probing the dynamic gating mechanism of other integral membrane proteins and ion channels.

MeSH Terms
Ion Channel Gating/genetics Mass Spectrometry/methods Models, Molecular Potassium Channels, Inwardly Rectifying/chemistry Protein Conformation Pulse Radiolysis Synchrotrons
Chemicals
Potassium Channels, Inwardly Rectifying
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Gupta Sayan
Center for Synchrotron Biosciences, Case Western Reserve University, Cleveland, OH 44022, USA.
Bavro Vassiliy N
D'Mello Rhijuta
Tucker Stephen J
Vénien-Bryan Catherine
Chance Mark R
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Article Info
Journal
Structure (London, England : 1993)
Abbr.
Structure
ISSN
1878-4186
DOI
10.1016/j.str.2010.04.012
Published
2010-07-14
Pages
839-46
Language
English
Region
United States
NLM ID
101087697
PMCID
PMC3124773
Subset
IM
Grants
British Heart Foundation · PG/09/016/26992 · United Kingdom
Biotechnology and Biological Sciences Research Council · BB/F013035/1 · United Kingdom
NIBIB NIH HHS · R01 EB009688 · United States
NIBIB NIH HHS · P30 EB009998 · United States
NIBIB NIH HHS · R01 EB009688-02 · United States
NIBIB NIH HHS · P30 EB009998-02 · United States
NIBIB NIH HHS · P41 EB001979 · United States
Wellcome Trust · 084655 · United Kingdom
External Links
PubMed source DOI: 10.1016/j.str.2010.04.012 PMC Full Text View journal details
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