Combining electron crystallography and X-ray crystallography to study the MlotiK1 cyclic nucleotide-regulated potassium channel

Gina M. Clayton*, Steve G. Aller, Jimin Wang, Vinzenz Unger, João H. Morais-Cabral

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

We have recently reported the X-ray structure of the cyclic nucleotide-regulated potassium channel, MlotiK1. Here we describe the application of both electron and X-ray crystallography to obtain high quality crystals. We suggest that the combined application of these techniques provides a useful strategy for membrane protein structure determination. We also present negative stain projection and cryo-data projection maps. These maps provide new insights about the properties of the MlotiK1 channel. In particular, a comparison of a 9 Å cryo-data projection with calculated model maps strongly suggests that there is a very weak interaction between the pore and the S1-S4 domains of this 6 TM tetrameric cation channel and that the S1-S4 domains can adopt multiple orientations relative to the pore.

Original languageEnglish (US)
Pages (from-to)220-226
Number of pages7
JournalJournal of Structural Biology
Volume167
Issue number3
DOIs
StatePublished - Sep 2009

Funding

The authors declare they have no competing financial interests. We would like to thank the staff of X25 and X29 beamlines at the National Synchrotron Light source and the Cornell High Energy Synchrotron Source (CHESS). This work was supported by grants from the American Heart Association (#0555822T), FCT (PTDC/QUI/66171/2006) and National Institutes of Health (GM068585) awarded to J.H.M.-C. and from PHS Grants GM66145, GM071590 awarded to V.M.U.

Keywords

  • 6 transmembrane helix channel
  • Crystallization conditions
  • Cyclic nucleotide-regulated potassium channel
  • Electron crystallography
  • Membrane protein
  • X-ray crystallography

ASJC Scopus subject areas

  • Structural Biology

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