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Title: Directly observed /sup 15/N NMR spectra of uniformly enriched proteins

Abstract

The proteins cytochrome c/sub 2/, cytochrome c', and ribulosebisphosphate carboxylase/oxygenase from Rhodospirillum rubrum were enriched in /sup 15/N by growth of the organism on /sup 15/NH/sub 4/Cl. The proteins were purified to homogeneity and studied by /sup 15/N NMR. Longitudinal and transverse relaxation times as well as the nuclear Overhauser effects were determined for various groups of the proteins which vary in molecular weight from 13,000 to 114,000. The values of these parameters for the amide resonances or for groups thought to be rigid were consistent with the molecular weights of the proteins. Relaxation times of the amino-terminal ..cap alpha..-amino groups and the side chain nitrogen atoms of arginine and lysine were consistent with much more rapid motion. Nitrogen atoms having bound protons were generally found to be decoupled from the protons by chemical exchange. Demonstrable /sup 1/H-/sup 15/N coupling was taken as an indication that exchange was hindered, either by hydrogen bonding interactions or by inaccessibility of the group to solvent. Histidine side chain nitrogen atoms, which experience a large chemical shift upon protonation/deprotonation, were often found to be broadened beyond detectability by chemical shift upon protonation/deprotonation, were often found to be broadened beyond detectability by chemical exchange andmore » tautomerization. Strategies for improving sensitivity and for obtaining specific peak assignments are also discussed.« less

Authors:
; ;
Publication Date:
Research Org.:
Univ. of California, Davis
OSTI Identifier:
5559509
Resource Type:
Journal Article
Journal Name:
Biochemistry; (United States)
Additional Journal Information:
Journal Volume: 26:8
Country of Publication:
United States
Language:
English
Subject:
62 RADIOLOGY AND NUCLEAR MEDICINE; CYTOCHROMES; NUCLEAR MAGNETIC RESONANCE; OXYGENASES; AMMONIUM CHLORIDES; CHEMICAL SHIFT; NITROGEN 15; NMR SPECTRA; OVERHAUSER EFFECT; PHOTOSYNTHESIS; PROTONS; RELAXATION TIME; RHODOSPIRILLUM; AMMONIUM COMPOUNDS; AMMONIUM HALIDES; BACTERIA; BARYONS; CHEMICAL REACTIONS; CHLORIDES; CHLORINE COMPOUNDS; ELEMENTARY PARTICLES; ENZYMES; FERMIONS; HADRONS; HALIDES; HALOGEN COMPOUNDS; ISOTOPES; LIGHT NUCLEI; MAGNETIC RESONANCE; MICROORGANISMS; NITROGEN ISOTOPES; NUCLEI; NUCLEONS; ODD-EVEN NUCLEI; ORGANIC COMPOUNDS; OXIDOREDUCTASES; PHOTOCHEMICAL REACTIONS; PIGMENTS; PROTEINS; RESONANCE; SPECTRA; STABLE ISOTOPES; SYNTHESIS; 550601* - Medicine- Unsealed Radionuclides in Diagnostics

Citation Formats

Smith, G M, Yu, L P, and Domingues, D J. Directly observed /sup 15/N NMR spectra of uniformly enriched proteins. United States: N. p., 1987. Web.
Smith, G M, Yu, L P, & Domingues, D J. Directly observed /sup 15/N NMR spectra of uniformly enriched proteins. United States.
Smith, G M, Yu, L P, and Domingues, D J. 1987. "Directly observed /sup 15/N NMR spectra of uniformly enriched proteins". United States.
@article{osti_5559509,
title = {Directly observed /sup 15/N NMR spectra of uniformly enriched proteins},
author = {Smith, G M and Yu, L P and Domingues, D J},
abstractNote = {The proteins cytochrome c/sub 2/, cytochrome c', and ribulosebisphosphate carboxylase/oxygenase from Rhodospirillum rubrum were enriched in /sup 15/N by growth of the organism on /sup 15/NH/sub 4/Cl. The proteins were purified to homogeneity and studied by /sup 15/N NMR. Longitudinal and transverse relaxation times as well as the nuclear Overhauser effects were determined for various groups of the proteins which vary in molecular weight from 13,000 to 114,000. The values of these parameters for the amide resonances or for groups thought to be rigid were consistent with the molecular weights of the proteins. Relaxation times of the amino-terminal ..cap alpha..-amino groups and the side chain nitrogen atoms of arginine and lysine were consistent with much more rapid motion. Nitrogen atoms having bound protons were generally found to be decoupled from the protons by chemical exchange. Demonstrable /sup 1/H-/sup 15/N coupling was taken as an indication that exchange was hindered, either by hydrogen bonding interactions or by inaccessibility of the group to solvent. Histidine side chain nitrogen atoms, which experience a large chemical shift upon protonation/deprotonation, were often found to be broadened beyond detectability by chemical shift upon protonation/deprotonation, were often found to be broadened beyond detectability by chemical exchange and tautomerization. Strategies for improving sensitivity and for obtaining specific peak assignments are also discussed.},
doi = {},
url = {https://www.osti.gov/biblio/5559509}, journal = {Biochemistry; (United States)},
number = ,
volume = 26:8,
place = {United States},
year = {Tue Apr 21 00:00:00 EDT 1987},
month = {Tue Apr 21 00:00:00 EDT 1987}
}