Enzymes: propionate CoA-transferase (EC 18.104.22.168); glutaconate CoA-transferase (EC 22.214.171.124).
Propionate CoA-transferase from Clostridium propionicum
Cloning of the gene and identification of glutamate 324 at the active site
Article first published online: 30 OCT 2003
European Journal of Biochemistry
Volume 269, Issue 1, pages 372–380, January 2002
How to Cite
Selmer, T., Willanzheimer, A. and Hetzel, M. (2002), Propionate CoA-transferase from Clostridium propionicum. European Journal of Biochemistry, 269: 372–380. doi: 10.1046/j.0014-2956.2001.02659.x
- Issue published online: 30 OCT 2003
- Article first published online: 30 OCT 2003
- (Received 3 August 2001, revised 5 November 2001, accepted 7 November 2001)
- Clostridium propionicum;
- alanine metabolism;
- active site;
- thiol ester
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.