Partial purification and characterization of cytidine 5'-diphosphate-diglyceride hydrolase from membranes of Escherichia coli

J Bacteriol. 1976 Mar;125(3):855-63. doi: 10.1128/jb.125.3.855-863.1976.

Abstract

Cytidine 5'-diphosphate (CDP)-diglyceride is hydrolyzed to phosphatidic acid and cytidine 5'-monophosphate by a specific membrane-bound enzyme in cell-free extracts of Escherichia coli. The hydrolase can be extracted from the particulate fraction with Triton X-100 and purified 1,000-fold in the presence of this detergent. Several nucleoside disphosphate diglycerides were synthesized to determine the substrate specificity of the hydrolase. CDP-diglyceride was hydrolyzed preferentially, although uridine 5'-diphosphate-diglyceride, guanosine 5'-diphosphate-diglyceride, and adenosine 5'-diphosphate (ADP)-diglyceride were also slowly hydrolyzed. Surprisingly, the purified enzyme did not catalyze detectable cleavage of deoxy-CDP (dCDP)-diglyceride. The liponucleotide pool of E. coli contains dCDP-diglyceride and CDP-diglyceride in approximately equal amounts (Raetz and Kennedy, 1973). Water-soluble nucleoside pyrophosphates, such as CDP-choline, nicotinamide adenine dinucleotide, or adenosine 5'-triphosphate are not attacked by this specific hydrolase. Hydrolysis of CDP-diglyceride is strongly inhibited by adenosine 5'-monophosphate and by ADP-diglyceride.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cell Membrane / enzymology
  • Cell-Free System
  • Chemical Precipitation
  • Cytidine Diphosphate Diglycerides / metabolism
  • Cytosine Nucleotides / biosynthesis
  • Hydrolases* / isolation & purification
  • Hydrolases* / metabolism
  • Hydrolysis
  • Phosphatidic Acids / biosynthesis
  • Polyethylene Glycols

Substances

  • Cytidine Diphosphate Diglycerides
  • Cytosine Nucleotides
  • Phosphatidic Acids
  • Polyethylene Glycols
  • Hydrolases