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J Struct Funct Genomics. 2009 Mar;10(1):9-16. doi: 10.1007/s10969-008-9046-7. Epub 2008 Nov 22.

Ratiocinative screen of eukaryotic integral membrane protein expression and solubilization for structure determination.

Author information

1
Department of Biochemistry and Biophysics, University of California at San Francisco, San Francisco, CA 94158-2517, USA. haysf@msg.ucsf.edu

Abstract

Persistent hurdles impede the successful determination of high-resolution crystal structures of eukaryotic integral membrane proteins (IMP). We designed a high-throughput structural genomics oriented pipeline that seeks to minimize effort in uncovering high-quality, responsive non-redundant targets for crystallization. This "discovery-oriented" pipeline sidesteps two significant bottlenecks in the IMP structure determination pipeline: expression and membrane extraction with detergent. In addition, proteins that enter the pipeline are then rapidly vetted by their presence in the included volume on a size-exclusion column--a hallmark of well-behaved IMP targets. A screen of 384 rationally selected eukaryotic IMPs in baker's yeast Saccharomyces cerevisiae is outlined to demonstrate the results expected when applying this discovery-oriented pipeline to whole-organism membrane proteomes.

PMID:
19031011
PMCID:
PMC2756966
DOI:
10.1007/s10969-008-9046-7
[Indexed for MEDLINE]
Free PMC Article

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