Open AccessPropionate CoA‐transferase from Clostridium propionicum
Author(s) -
Selmer Thorsten,
Willanzheimer Angela,
Hetzel Marc
Publication year - 2002
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1046/j.0014-2956.2001.02659.x
Subject(s) - transferase , peptidyl transferase , propionate , biochemistry , enzyme , active site , escherichia coli , hydroxylamine , chemistry , stereochemistry , biology , gene , rna , ribosome
Propionate CoA‐transferase from Clostridium propionicum has been purified and the gene encoding the enzyme has been cloned and sequenced. The enzyme was rapidly and irreversibly inactivated by sodium borohydride or hydroxylamine in the presence of propionyl‐CoA. The reduction of the thiol ester between a catalytic site glutamate and CoA with borohydride and the cleavage by hydroxylamine were used to introduce a site‐specific label, which was followed by␣MALDI‐TOF‐MS. This allowed the identification of glutamate 324 at the active site. Propionate CoA‐transferase and similar proteins deduced from the genomes of Escherichia coli , Staphylococcus aureus , Bacillus halodurans and Aeropyrum pernix are proposed to form a novel subclass of CoA‐transferases. Secondary structure element predictions were generated and compared to known crystal structures in the databases. A high degree of structural similarity was observed between the arrangement of secondary structure elements in these proteins and glutaconate CoA‐transferase from Acidaminococcus fermentans .