Electron spin-echo envelope modulation (ESEEM) reveals water and phosphate interactions with the KcsA potassium channel

John A. Cieslak, Pamela J. Focia, Adrian Gross*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

33 Scopus citations

Abstract

Electron spin-echo envelope modulation (ESEEM) spectroscopy is a well-established technique for the study of naturally occurring paramagnetic metal centers. The technique has been used to study copper complexes, hemes, enzyme mechanisms, micellar water content, and water permeation profiles in membranes, among other applications. In the present study, we combine ESEEM spectroscopy with site-directed spin labeling (SDSL) and X-ray crystallography in order to evaluate the technique's potential as a structural tool to describe the native environment of membrane proteins. Using the KcsA potassium channel as a model system, we demonstrate that deuterium ESEEM can detect water permeation along the lipid-exposed surface of the KcsA outer helix. We further demonstrate that 31P ESEEM is able to identify channel residues that interact with the phosphate headgroup of the lipid bilayer. In combination with X-ray crystallography, the 31P data may be used to define the phosphate interaction surface of the protein. The results presented here establish ESEEM as a highly informative technique for SDSL studies of membrane proteins.

Original languageEnglish (US)
Pages (from-to)1486-1494
Number of pages9
JournalBiochemistry
Volume49
Issue number7
DOIs
StatePublished - Feb 23 2010

ASJC Scopus subject areas

  • Biochemistry

Fingerprint Dive into the research topics of 'Electron spin-echo envelope modulation (ESEEM) reveals water and phosphate interactions with the KcsA potassium channel'. Together they form a unique fingerprint.

Cite this