title: Ratiocinative screen of eukaryotic integral membrane protein expression and solubilization for structure determination.
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.
authors.affiliation: Department of Biochemistry and Biophysics, University of California at San Francisco, San Francisco, CA 94158-2517, USA. haysf@msg.ucsf.edu
authors.forename: Franklin A
authors.initials: FA
authors.lastname: Hays
authors.forename: Zygy
authors.initials: Z
authors.lastname: Roe-Zurz
authors.forename: Min
authors.initials: M
authors.lastname: Li
authors.forename: Libusha
authors.initials: L
authors.lastname: Kelly
authors.forename: Franz
authors.initials: F
authors.lastname: Gruswitz
authors.forename: Andrej
authors.initials: A
authors.lastname: Sali
authors.forename: Robert M
authors.initials: RM
authors.lastname: Stroud
mesh_terms: D000818:Animals; D018360:Crystallography, X-Ray; D030562:Databases, Protein; D023281:Genomics; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry*; D011487:Protein Conformation; D020543:Proteome / Q000032:analysis; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism
chemical_list: D008565:Membrane Proteins; D020543:Proteome; D029701:Saccharomyces cerevisiae Proteins