title: Solution structure of a soluble fragment derived from a membrane protein by shotgun proteolysis.
abstract: We have previously reported a phage display method for the identification of protein domains on a genome-wide scale (shotgun proteolysis). Here we present the solution structure of a fragment of the Escherichia coli membrane protein yrfF, as identified by shotgun proteolysis, and determined by NMR spectroscopy. Despite the absence of computational predictions, the fragment formed a well-defined beta-barrel structure, distantly falling within the OB-fold classification. Our results highlight the potential of high-throughput experimental approaches for the identification of protein domains for structural studies.
authors: MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH, UK.
authors: Mark D
authors: MD
authors: Allen
authors: Department of Immunology, Garvan Institute of Medical Research, 384 Victoria Street, Darlinghurst, Sydney, NSW 2010, Australia Faculty of Medicine, St Vincent's Clinical School, The University of New South Wales, Darlinghurst, Sydney, NSW 2010, Australia.
authors: Mary
authors: M
authors: Christie
authors: MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH, UK.
authors: Peter
authors: P
authors: Jones
authors: Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria 3800, Australia.
authors: Benjamin T
authors: BT
authors: Porebski
authors: Department of Immunology, Garvan Institute of Medical Research, 384 Victoria Street, Darlinghurst, Sydney, NSW 2010, Australia Faculty of Medicine, St Vincent's Clinical School, The University of New South Wales, Darlinghurst, Sydney, NSW 2010, Australia.
authors: Brendan
authors: B
authors: Roome
authors: MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH, UK.
authors: Stefan M V
authors: SM
authors: Freund
authors: Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria 3800, Australia.
authors: Ashley M
authors: AM
authors: Buckle
authors: MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH, UK.
authors: Mark
authors: M
authors: Bycroft
authors: Department of Immunology, Garvan Institute of Medical Research, 384 Victoria Street, Darlinghurst, Sydney, NSW 2010, Australia Faculty of Medicine, St Vincent's Clinical School, The University of New South Wales, Darlinghurst, Sydney, NSW 2010, Australia d.christ@garvan.org.au.
authors: Daniel
authors: D
authors: Christ
mesh_terms: D000595:Amino Acid Sequence; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism*; D017433:Protein Structure, Secondary; D059748:Proteolysis; D017421:Sequence Analysis; D012995:Solubility
chemical_list: D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D010446:Peptide Fragments; C000609030:yrfF protein, E coli
keywords: NMR spectroscopy; membrane proteins; phage display; protein domains; proteolysis