title: NMR for structural proteomics of Thermotoga maritima: screening and structure determination.
abstract: This paper describes the NMR screening of 141 small (<15 kDa) recombinant Thermotoga maritima proteins for globular folding. The experimental data shows that approximately 25% of the screened proteins are folded under our screening conditions, which makes this procedure an important step for selecting those proteins that are suitable for structure determination. A comparison of screening based either on 1D 1H NMR with unlabeled proteins or on 2D [1H,15N]-COSY with uniformly 15N-labeled proteins is presented, and a comprehensive analysis of the 1D 1H NMR screening data is described. As an illustration of the utility of these methods to structural proteomics, the NMR structure determination of TM1492 (ribosomal protein L29) is presented. This 66-residue protein consists of a N-terminal 3(10)-helix and two long alpha-helices connected by a tight turn centered about glycine 35, where conserved leucine and isoleucine residues in the two alpha-helices form a small hydrophobic core.
authors.affiliation: The Scripps Research Institute, Department of Molecular Biology and Joint Center of Structural Genomics, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA. wolfpeti@scripps.edu
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Peti
authors.forename: Touraj
authors.initials: T
authors.lastname: Etezady-Esfarjani
authors.forename: Torsten
authors.initials: T
authors.lastname: Herrmann
authors.forename: Heath E
authors.initials: HE
authors.lastname: Klock
authors.forename: Scott A
authors.initials: SA
authors.lastname: Lesley
authors.forename: Kurt
authors.initials: K
authors.lastname: Wüthrich
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D019906:Nuclear Magnetic Resonance, Biomolecular; D017434:Protein Structure, Tertiary; D040901:Proteomics / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D020124:Thermotoga maritima / Q000737:chemistry / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins