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Modification of bovine heart succinate dehydrogenase with ethoxyformic anhydride and Rose bengal: Evidence for essential histidyl residues protectable by substrates

Hederstedt, Lars LU and Hatefi, Youssef (1986) In Archives of Biochemistry and Biophysics 247(2). p.346-354
Abstract
Purified and membrane-bound succinate dehydrogenase (SDH) from bovine heart mitochondria was inhibited by the histidine-modifying reagents ethoxyformic anhydride (EFA) and Rose Bengal in the presence of light. Succinate and competitive inhibitors protected against inhibition, and decreased the number of histidyl residues modified by EFA. The essential residue modified by EFA was not the essential thiol of SDH, but modification of the essential thiol abolished the protective effect of malonate against inhibition of SDH by EFA. The EFA inhibition was reversed by hydroxylamine nearly completely when the inhibition was ⩽35%, and only partially when the inhibition was more extensive. The uv spectrum of EFA-modified SDH before and after... (More)
Purified and membrane-bound succinate dehydrogenase (SDH) from bovine heart mitochondria was inhibited by the histidine-modifying reagents ethoxyformic anhydride (EFA) and Rose Bengal in the presence of light. Succinate and competitive inhibitors protected against inhibition, and decreased the number of histidyl residues modified by EFA. The essential residue modified by EFA was not the essential thiol of SDH, but modification of the essential thiol abolished the protective effect of malonate against inhibition of SDH by EFA. The EFA inhibition was reversed by hydroxylamine nearly completely when the inhibition was ⩽35%, and only partially when the inhibition was more extensive. The uv spectrum of EFA-modified SDH before and after hydroxylamine treatment suggested that extensive inhibition of SDH with EFA may result in ethoxyformylation at both imidazole nitrogens of histidyl residues. Such a modification is not reversed by hydroxylamine. Succinate dehydrogenases and fumarate reductases from several different sources have similar compositions, and the two enzymes from Escherichia coli have considerable homology in the amino acid composition of their respective flavoprotein and iron-sulfur protein subunits. In the former, there is a short stretch containing conserved histidine, cysteine, and arginine residues. These residues, if also conserved in the bovine enzyme, may be the essential active site residues suggested by this work (histidine) and previously (cysteine, arginine). (Less)
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author
publishing date
type
Contribution to journal
publication status
published
subject
in
Archives of Biochemistry and Biophysics
volume
247
issue
2
pages
346 - 354
publisher
Academic Press
external identifiers
  • scopus:0022497345
ISSN
0003-9861
DOI
10.1016/0003-9861(86)90593-X
language
English
LU publication?
no
id
5b5b2e6c-2389-460f-8ddb-99aa64622a07
date added to LUP
2017-07-18 11:08:13
date last changed
2017-09-26 15:06:09
@article{5b5b2e6c-2389-460f-8ddb-99aa64622a07,
  abstract     = {Purified and membrane-bound succinate dehydrogenase (SDH) from bovine heart mitochondria was inhibited by the histidine-modifying reagents ethoxyformic anhydride (EFA) and Rose Bengal in the presence of light. Succinate and competitive inhibitors protected against inhibition, and decreased the number of histidyl residues modified by EFA. The essential residue modified by EFA was not the essential thiol of SDH, but modification of the essential thiol abolished the protective effect of malonate against inhibition of SDH by EFA. The EFA inhibition was reversed by hydroxylamine nearly completely when the inhibition was ⩽35%, and only partially when the inhibition was more extensive. The uv spectrum of EFA-modified SDH before and after hydroxylamine treatment suggested that extensive inhibition of SDH with EFA may result in ethoxyformylation at both imidazole nitrogens of histidyl residues. Such a modification is not reversed by hydroxylamine. Succinate dehydrogenases and fumarate reductases from several different sources have similar compositions, and the two enzymes from Escherichia coli have considerable homology in the amino acid composition of their respective flavoprotein and iron-sulfur protein subunits. In the former, there is a short stretch containing conserved histidine, cysteine, and arginine residues. These residues, if also conserved in the bovine enzyme, may be the essential active site residues suggested by this work (histidine) and previously (cysteine, arginine).},
  author       = {Hederstedt, Lars and Hatefi, Youssef},
  issn         = {0003-9861},
  language     = {eng},
  number       = {2},
  pages        = {346--354},
  publisher    = {Academic Press},
  series       = {Archives of Biochemistry and Biophysics},
  title        = {Modification of bovine heart succinate dehydrogenase with ethoxyformic anhydride and Rose bengal: Evidence for essential histidyl residues protectable by substrates},
  url          = {http://dx.doi.org/10.1016/0003-9861(86)90593-X},
  volume       = {247},
  year         = {1986},
}