sup 1 H, sup 15 N, and sup 13 C NMR signal assignments of III sup Glc , a signal-transducing protein of Escherichia coli, using three-dimensional triple-resonance techniques

• Published in: Biochemistry (Easton) Vol. 30:41
• Main Authors: Pelton, J.G., Torchia, D.A., Meadow, N.D., Cingyuen Wong, Roseman, S.
• Format: Journal Article
• Language: English
• Published: United States 15.10.1991
• Subjects: 550601 - Medicine- Unsealed Radionuclides in Diagnostics; BACTERIA; BARYONS; CARBON 13; CARBON ISOTOPES; CHEMICAL SHIFT; ELEMENTARY PARTICLES; ENZYMES; ESCHERICHIA COLI; EVEN-ODD NUCLEI; FERMIONS; HADRONS; ISOTOPES; LIGHT NUCLEI; MAGNETIC RESONANCE; MEMBRANE PROTEINS; MEMBRANE TRANSPORT; MICROORGANISMS; NITROGEN 15; NITROGEN ISOTOPES; NUCLEAR MAGNETIC RESONANCE; NUCLEI; NUCLEONS; ODD-EVEN NUCLEI; ORGANIC COMPOUNDS; PHOSPHOENOLPYRUVATE; PHOSPHORUS-GROUP TRANSFERASES; PHOSPHOTRANSFERASES; PROTEINS; PROTONS; RADIOLOGY AND NUCLEAR MEDICINE; RESONANCE; STABLE ISOTOPES; TRANSFERASES
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi00105a032

III{sup Glc} is an 18.1-kDa signal-transducing phosphocarrier protein of the phosphoenolpyruvate:glycose phosphotransferase system (PTS) of Escherichia coli. Virtually complete (98%) backbone {sup 1}H, {sup 15}N, and {sup 13}C nuclear magnetic resonance (NMR) signal assignments were determined by using a battery of triple-resonance three-dimensional (3D) NMR pulse sequences. In addition, nearly complete side-chain {sup 1}H and {sup 13}C signal assignments were obtained from an analysis of 3D {sup 13}C HCCH-COSY and HCCH-TOCSY spectra. These experiments rely almost exclusively upon one- and two-bond J couplings to transfer magnetization and to correlate proton and heteronuclear NMR signals. Hence, essentially complete signal assignments of this 168-residue protein were made without any assumptions regarding secondary structure and without the aid of a crystal structure, which is not yet available. Moreover, only three samples, one uniformly {sup 15}N-enriched, one uniformly {sup 15}N/{sup 13}C-enriched, and one containing a few types of amino acids labeled with {sup 15}N and/or {sup 13}C, were needed to make the assignments. The nearly complete set of backbone and side-chain atom assignments reported herein provide a basis for studies of the three-dimensional structure and dynamics of III{sup Glc} as well as its interactions with a variety of membrane and cytoplasmic proteins.