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

Franklin A. Hays, Zygy Roe-Zurz, Min Li, Libusha Kelly, Franz Gruswitz, Andrej Sali, Robert M. Stroud

Research output: Contribution to journalReview articlepeer-review

9 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)9-16
Number of pages8
JournalJournal of Structural and Functional Genomics
Volume10
Issue number1
DOIs
StatePublished - Mar 1 2009
Externally publishedYes

Keywords

  • Discovery-oriented screen
  • Eukaryotic integral membrane protein
  • Membrane protein structure
  • Saccharomyces cerevisiae
  • Structural genomics

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry
  • Genetics

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