Phosphopantothenoylcysteine synthetase from Escherichia coli. Identification and characterization of the last unidentified coenzyme A biosynthetic enzyme in bacteria

J Biol Chem. 2001 Apr 27;276(17):13513-6. doi: 10.1074/jbc.C100033200. Epub 2001 Mar 13.

Abstract

Phosphopantothenoylcysteine synthase catalyzes the formation of (R)-4'-phospho-N-pantothenoylcysteine from 4'-phosphopantothenate and l-cysteine: this enzyme, involved in the biosynthesis of coenzyme A (CoA), has not previously been identified. Recently it was shown that the NH(2)-terminal domain of the Dfp protein from bacteria catalyzes the next step in CoA biosynthesis, the decarboxylation of (R)-4'-phospho-N-pantothenoylcysteine to form 4'-phosphopantetheine (Kupke, T., Uebele, M., Schmid, D., Jung, G., Blaesse, M., and Steinbacher, S. (2000) J. Biol. Chem. 275, 31838-31846). We have partially purified phosphopantothenoylcysteine decarboxylase from Escherichia coli and demonstrated that the protein encoded by the dfp gene, here renamed coaBC, also has phosphopantothenoylcysteine synthetase activity, using CTP rather than ATP as the activating nucleoside 5'-triphosphate. This discovery completes the identification of all the enzymes involved in the biosynthesis of coenzyme A in bacteria.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Carboxy-Lyases / chemistry*
  • Carboxy-Lyases / genetics
  • Carboxy-Lyases / metabolism
  • Cloning, Molecular
  • Coenzyme A / biosynthesis*
  • Cysteine / analogs & derivatives
  • Cysteine / metabolism
  • Cytidine Monophosphate / metabolism
  • Cytidine Triphosphate / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / enzymology*
  • Flavins / metabolism
  • Kinetics
  • Models, Chemical
  • Pantothenic Acid / analogs & derivatives
  • Pantothenic Acid / chemistry
  • Pantothenic Acid / metabolism
  • Peptide Synthases / chemistry*
  • Peptide Synthases / genetics
  • Peptide Synthases / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Spectrometry, Mass, Electrospray Ionization
  • Ultraviolet Rays

Substances

  • Flavins
  • Recombinant Proteins
  • Pantothenic Acid
  • Cytidine Triphosphate
  • pantothenylcysteine 4'-phosphate
  • Adenosine Triphosphate
  • Carboxy-Lyases
  • Phosphopantothenoyl-cysteine decarboxylase
  • Peptide Synthases
  • phosphopantothenoyl-cysteine synthetase
  • Cytidine Monophosphate
  • Cysteine
  • Coenzyme A