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

Journal Article · Tue Oct 15 00:00:00 EDT 1991 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00105a032· OSTI ID:5713515

Pelton, J G; Torchia, D A ^[1]; Meadow, N D; Wong, Cingyuen; Roseman, S ^[2]

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1. National Inst. of Health, Bethesda, MD (United States)
2. Johns Hopkins Univ., Baltimore, MD (United States)

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.

• OSTI ID: 5713515
• Journal Information: Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 30:41; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550601* -- Medicine-- Unsealed Radionuclides in Diagnostics
62 RADIOLOGY AND NUCLEAR MEDICINE
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
RESONANCE
STABLE ISOTOPES
TRANSFERASES