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Title: Proton resonance assignments of horse ferricytochrome c

Abstract

Two-dimensional nuclear magnetic resonance spectroscopy (2D NMR) was used to obtain extensive resonance assignments in the {sup 1}H NMR spectrum of horse ferricytochrome c. Assignments were made for the main-chain and C{sub {beta}} protons of 102 residues (all except Pro-44 and Gly-84) and the majority of side-chain protons. As starting points for the assignment of the oxidized protein, a limited set of protons was initially assigned by use of 2D NMR magnetization transfer methods to correlate resonances in the oxidized form with assigned resonances in the reduced form. Given the complexity of the spectrum due to the size of this protein (104 residues) and its paramagnetic center, the initial search for side-chain spin systems in J-correlated spectra was successful only for the simplest side chains, but the majority of NH-C{sub {alpha}}H-C{sub {beta}}H subspin systems (NAB sets) could be identified at this stage. The subsequent search for sequential NOE connectivities focused on NAB sets, with use of previously assigned residues to place NOE-connected segments within the amino acid sequence. Selective proton labeling of either the slowly or the rapidly exchanging amide sites was used to simplify the spectra, and systematic work at two temperatures was used to resolve ambiguities in themore » 2D NMR spectra. These approaches, together with the use of magnetization transfer methods to correlate reduced and oxidized cytochrome c spectra, provide multiple cross-checks to verify assignments.« less

Authors:
; ; ; ;  [1]
  1. Univ. of Pennsylvania, Philadelphia (USA)
Publication Date:
OSTI Identifier:
5640791
Resource Type:
Journal Article
Journal Name:
Biochemistry; (USA)
Additional Journal Information:
Journal Volume: 28:1; Journal ID: ISSN 0006-2960
Country of Publication:
United States
Language:
English
Subject:
62 RADIOLOGY AND NUCLEAR MEDICINE; CYTOCHROMES; NUCLEAR MAGNETIC RESONANCE; STRUCTURE-ACTIVITY RELATIONSHIPS; CRYSTALLOGRAPHY; HORSES; IRON COMPOUNDS; PROTONS; ANIMALS; BARYONS; ELEMENTARY PARTICLES; FERMIONS; HADRONS; MAGNETIC RESONANCE; MAMMALS; NUCLEONS; ORGANIC COMPOUNDS; PIGMENTS; PROTEINS; RESONANCE; TRANSITION ELEMENT COMPOUNDS; VERTEBRATES; 550601* - Medicine- Unsealed Radionuclides in Diagnostics

Citation Formats

Feng, Y, Roder, H, Englander, S W, Wand, A J, and Di Stefano, D L. Proton resonance assignments of horse ferricytochrome c. United States: N. p., 1989. Web. doi:10.1021/bi00427a027.
Feng, Y, Roder, H, Englander, S W, Wand, A J, & Di Stefano, D L. Proton resonance assignments of horse ferricytochrome c. United States. https://doi.org/10.1021/bi00427a027
Feng, Y, Roder, H, Englander, S W, Wand, A J, and Di Stefano, D L. 1989. "Proton resonance assignments of horse ferricytochrome c". United States. https://doi.org/10.1021/bi00427a027.
@article{osti_5640791,
title = {Proton resonance assignments of horse ferricytochrome c},
author = {Feng, Y and Roder, H and Englander, S W and Wand, A J and Di Stefano, D L},
abstractNote = {Two-dimensional nuclear magnetic resonance spectroscopy (2D NMR) was used to obtain extensive resonance assignments in the {sup 1}H NMR spectrum of horse ferricytochrome c. Assignments were made for the main-chain and C{sub {beta}} protons of 102 residues (all except Pro-44 and Gly-84) and the majority of side-chain protons. As starting points for the assignment of the oxidized protein, a limited set of protons was initially assigned by use of 2D NMR magnetization transfer methods to correlate resonances in the oxidized form with assigned resonances in the reduced form. Given the complexity of the spectrum due to the size of this protein (104 residues) and its paramagnetic center, the initial search for side-chain spin systems in J-correlated spectra was successful only for the simplest side chains, but the majority of NH-C{sub {alpha}}H-C{sub {beta}}H subspin systems (NAB sets) could be identified at this stage. The subsequent search for sequential NOE connectivities focused on NAB sets, with use of previously assigned residues to place NOE-connected segments within the amino acid sequence. Selective proton labeling of either the slowly or the rapidly exchanging amide sites was used to simplify the spectra, and systematic work at two temperatures was used to resolve ambiguities in the 2D NMR spectra. These approaches, together with the use of magnetization transfer methods to correlate reduced and oxidized cytochrome c spectra, provide multiple cross-checks to verify assignments.},
doi = {10.1021/bi00427a027},
url = {https://www.osti.gov/biblio/5640791}, journal = {Biochemistry; (USA)},
issn = {0006-2960},
number = ,
volume = 28:1,
place = {United States},
year = {Tue Jan 10 00:00:00 EST 1989},
month = {Tue Jan 10 00:00:00 EST 1989}
}