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New properties of Bacillus subtilis succinate dehydrogenase altered at the active site

Hederstedt, Lars LU and Hedén, Lars-Olof LU (1989) In Biochemical Journal 260(2). p.491-497
Abstract
Mammalian and Escherichia coli succinate dehydrogenase (SDH) and E. coli fumarate reductase apparently
contain an essential cysteine residue at the active site, as shown by substrate-protectable inactivation with
thiol-specific reagents. Bacillus subtilis SDH was found to be resistant to this type of reagent and contains
an alanine residue at the amino acid position equivalent to the only invariant cysteine in the flavoprotein
subunit of E. coli succinate oxidoreductases. Substitution of this alanine, at position 252 in the flavoprotein subunit of B. subtilis SDH, by cysteine resulted in an enzyme sensitive to thiol-specific reagents and protectable by substrate. Other biochemical properties of the redesigned SDH were... (More)
Mammalian and Escherichia coli succinate dehydrogenase (SDH) and E. coli fumarate reductase apparently
contain an essential cysteine residue at the active site, as shown by substrate-protectable inactivation with
thiol-specific reagents. Bacillus subtilis SDH was found to be resistant to this type of reagent and contains
an alanine residue at the amino acid position equivalent to the only invariant cysteine in the flavoprotein
subunit of E. coli succinate oxidoreductases. Substitution of this alanine, at position 252 in the flavoprotein subunit of B. subtilis SDH, by cysteine resulted in an enzyme sensitive to thiol-specific reagents and protectable by substrate. Other biochemical properties of the redesigned SDH were similar to those of the wild-type enzyme. It is concluded that the invariant cysteine in the flavoprotein of E. coli succinate oxidoreductases corresponds to the active site thiol. However, this cysteine is most likely not essential for succinate oxidation and seemingly lacks an assignable specific function. An invariant arginine in juxtaposition to Ala-252 in the flavoprotein of B. subtilis SDH, and to the invariant cysteine in the E. coli homologous enzymes, is probably essential for substrate binding. (Less)
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author
organization
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Contribution to journal
publication status
published
subject
in
Biochemical Journal
volume
260
issue
2
pages
491 - 497
publisher
Portland Press Limited
ISSN
0264-6021
DOI
10.1042/bj2600491
language
English
LU publication?
yes
id
68640c32-f9f3-447e-b006-a0653f22d0b1
date added to LUP
2017-07-18 10:50:06
date last changed
2017-09-07 11:44:47
@article{68640c32-f9f3-447e-b006-a0653f22d0b1,
  abstract     = {Mammalian and Escherichia coli succinate dehydrogenase (SDH) and E. coli fumarate reductase apparently<br/>contain an essential cysteine residue at the active site, as shown by substrate-protectable inactivation with<br/>thiol-specific reagents. Bacillus subtilis SDH was found to be resistant to this type of reagent and contains<br/>an alanine residue at the amino acid position equivalent to the only invariant cysteine in the flavoprotein<br/>subunit of E. coli succinate oxidoreductases. Substitution of this alanine, at position 252 in the flavoprotein subunit of B. subtilis SDH, by cysteine resulted in an enzyme sensitive to thiol-specific reagents and protectable by substrate. Other biochemical properties of the redesigned SDH were similar to those of the wild-type enzyme. It is concluded that the invariant cysteine in the flavoprotein of E. coli succinate oxidoreductases corresponds to the active site thiol. However, this cysteine is most likely not essential for succinate oxidation and seemingly lacks an assignable specific function. An invariant arginine in juxtaposition to Ala-252 in the flavoprotein of B. subtilis SDH, and to the invariant cysteine in the E. coli homologous enzymes, is probably essential for substrate binding.},
  author       = {Hederstedt, Lars and Hedén, Lars-Olof},
  issn         = {0264-6021},
  language     = {eng},
  number       = {2},
  pages        = {491--497},
  publisher    = {Portland Press Limited},
  series       = {Biochemical Journal},
  title        = {New properties of <em>Bacillus subtilis</em> succinate dehydrogenase altered at the active site},
  url          = {http://dx.doi.org/10.1042/bj2600491},
  volume       = {260},
  year         = {1989},
}