title: Sequence-specific NMR assignments of the trp repressor from Escherichia coli using three-dimensional 15N/1H heteronuclear techniques.
abstract: Sequence-specific 15N and 1H assignments for the trp holorepressor from Escherichia coli are reported. The trp repressor consists of two identical 107-residue subunits which are highly helical in the crystal state [Schevitz, R., Otwinowski, Z., Joachimiak, A., Lawson, C. L. & Sigler, P. B. (1985) Nature 317, 782-786]. The high helical content and the relatively large size of the protein (Mr = 25,000) make it difficult to assign even the main-chain resonances by conventional homonuclear two-dimensional NMR methods. However, we have now assigned the main-chain resonances of 94% of the residues by using three-dimensional 15N/1H heteronuclear experiments on a sample of protein uniformly labelled with 15N. The additional resolution obtained by spreading out the signals into three dimensions proved indispensable in making these assignments. In particular, we have been able to resolve signals from residues in the N-terminal region of the A helix for the first time in solution. The observed NOE results confirm that the repressor is highly helical in solution, and contains no extended chain conformations.
authors.affiliation: Laboratory of Molecular Structure, National Institute for Medical Research, London, England.
authors.forename: K L
authors.initials: KL
authors.lastname: Borden
authors.forename: C J
authors.initials: CJ
authors.lastname: Bauer
authors.forename: T A
authors.initials: TA
authors.lastname: Frenkiel
authors.forename: P
authors.initials: P
authors.lastname: Beckmann
authors.forename: A N
authors.initials: AN
authors.lastname: Lane
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D004926:Escherichia coli / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009587:Nitrogen Isotopes; D012097:Repressor Proteins / Q000737:chemistry*; D029968:Escherichia coli Proteins
chemical_list: D001426:Bacterial Proteins; D009587:Nitrogen Isotopes; D012097:Repressor Proteins; C027190:TRPR protein, E coli; D029968:Escherichia coli Proteins