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Title: Proton NMR assignments of heme contracts and catalytically implicated amino acids in cyanide-ligated cytochrome c peroxidase determined from one- and two-dimensional nuclear Overhauser effects

Abstract

Proton NMR assignments of the heme pocket and catalytically relevant amino acid protons have been accomplished for cyanide-ligated yeast cytochrome c peroxidase. This form of the protein, while not enzymatically active itself, is the best model available (that displays a resolvable proton NMR spectrum) for the six-coordinate low-spin active intermediates, compounds I and II. The assignments were made with a combination of one- and two-dimensional nuclear Overhauser effect methods and demonstrate the utility of NOESY experiments for paramagnetic proteins of relatively large size (M{sub r} 34,000). Assignments of both isotope exchangeable and nonexchangeable proton resonances were obtained by using enzyme preparations in both 90% H{sub 2}O/10% D{sub 2}O and, separately, in 99.9% D{sub 2}O solvent systems. Complete resonance assignments have been achieved for the proximal histidine, His-175, and His-52, which is a member of the catalytic triad on the distal side of the heme. In addition, partial assignments are reported for Trp-51 and Arg-48, catalytically important residues, both on the distal side. Aside from His-175, partial assignments for amino acids on the proximal side of the heme are proposed for the alanines at primary sequence positions 174 and 176 and for Thr-180 and Leu-232.

Authors:
 [1];  [2]
  1. (Washington State Univ., Pullman (USA))
  2. (Northern Illinois Univ., DeKalb (USA))
Publication Date:
OSTI Identifier:
5594898
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemistry; (United States); Journal Volume: 30:18
Country of Publication:
United States
Language:
English
Subject:
62 RADIOLOGY AND NUCLEAR MEDICINE; AMINO ACIDS; NUCLEAR MAGNETIC RESONANCE; HEME; PEROXIDASES; MOLECULAR STRUCTURE; ARGININE; CYTOCHROME OXIDASE; CYTOCHROMES; HEAVY WATER; HISTIDINE; MOLECULAR WEIGHT; OVERHAUSER EFFECT; PROTONS; TRYPTOPHAN; YEASTS; AROMATICS; AZAARENES; AZOLES; BARYONS; CARBOXYLIC ACIDS; ELEMENTARY PARTICLES; ENZYMES; EUMYCOTA; FERMIONS; FUNGI; HADRONS; HAEM DEHYDROGENASES; HETEROCYCLIC ACIDS; HETEROCYCLIC COMPOUNDS; HYDROGEN COMPOUNDS; IMIDAZOLES; INDOLES; MAGNETIC RESONANCE; MICROORGANISMS; NUCLEONS; ORGANIC ACIDS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; OXIDOREDUCTASES; OXYGEN COMPOUNDS; PIGMENTS; PLANTS; PORPHYRINS; PROTEINS; PYRROLES; RESONANCE; WATER; 550601* - Medicine- Unsealed Radionuclides in Diagnostics

Citation Formats

Satterlee, J.D., and Erman, J.E. Proton NMR assignments of heme contracts and catalytically implicated amino acids in cyanide-ligated cytochrome c peroxidase determined from one- and two-dimensional nuclear Overhauser effects. United States: N. p., 1991. Web. doi:10.1021/bi00232a005.
Satterlee, J.D., & Erman, J.E. Proton NMR assignments of heme contracts and catalytically implicated amino acids in cyanide-ligated cytochrome c peroxidase determined from one- and two-dimensional nuclear Overhauser effects. United States. doi:10.1021/bi00232a005.
Satterlee, J.D., and Erman, J.E. Tue . "Proton NMR assignments of heme contracts and catalytically implicated amino acids in cyanide-ligated cytochrome c peroxidase determined from one- and two-dimensional nuclear Overhauser effects". United States. doi:10.1021/bi00232a005.
@article{osti_5594898,
title = {Proton NMR assignments of heme contracts and catalytically implicated amino acids in cyanide-ligated cytochrome c peroxidase determined from one- and two-dimensional nuclear Overhauser effects},
author = {Satterlee, J.D. and Erman, J.E.},
abstractNote = {Proton NMR assignments of the heme pocket and catalytically relevant amino acid protons have been accomplished for cyanide-ligated yeast cytochrome c peroxidase. This form of the protein, while not enzymatically active itself, is the best model available (that displays a resolvable proton NMR spectrum) for the six-coordinate low-spin active intermediates, compounds I and II. The assignments were made with a combination of one- and two-dimensional nuclear Overhauser effect methods and demonstrate the utility of NOESY experiments for paramagnetic proteins of relatively large size (M{sub r} 34,000). Assignments of both isotope exchangeable and nonexchangeable proton resonances were obtained by using enzyme preparations in both 90% H{sub 2}O/10% D{sub 2}O and, separately, in 99.9% D{sub 2}O solvent systems. Complete resonance assignments have been achieved for the proximal histidine, His-175, and His-52, which is a member of the catalytic triad on the distal side of the heme. In addition, partial assignments are reported for Trp-51 and Arg-48, catalytically important residues, both on the distal side. Aside from His-175, partial assignments for amino acids on the proximal side of the heme are proposed for the alanines at primary sequence positions 174 and 176 and for Thr-180 and Leu-232.},
doi = {10.1021/bi00232a005},
journal = {Biochemistry; (United States)},
number = ,
volume = 30:18,
place = {United States},
year = {Tue May 07 00:00:00 EDT 1991},
month = {Tue May 07 00:00:00 EDT 1991}
}