Lipid-dependent membrane enzymes. Purification to homogeneity and further characterization of diacylglycerol kinase from Escherichia coli

Eur J Biochem. 1988 Jan 15;171(1-2):335-42. doi: 10.1111/j.1432-1033.1988.tb13795.x.


1. Diacylglycerol kinase apoprotein was purified from membranes of Escherichia coli K12 by a six-step procedure that included HPLC. The proposed assignment of the enzyme to the dgkA gene [Lightner et al. (1983) J. Biol. Chem. 258, 10856-10861] could be supported by molecular mass determination (approximately 14 kDa), N-terminal sequencing (Met-Ala-Asn), cyanogen bromide fragmentation and amino acid analysis. As predicted, proline was absent. 2. The membrane-associated as well as the butan-1-ol-dissolved enzyme survived heating to 100 degrees C. 3. Alkylglycoside detergents were found to constitute an additional class of lipid activators. 4. The enzyme apoprotein in a non-activating substrate/detergent solution was capable of autocatalytic self-activation which was attributed to a novel feedback activation mechanism involving phosphatidic acid (diacylglycerol 3-phosphate).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amino Acids / analysis
  • Cell Membrane / enzymology
  • Cyanogen Bromide
  • Detergents / pharmacology
  • Diacylglycerol Kinase
  • Enzyme Activation
  • Escherichia coli / enzymology*
  • Hot Temperature
  • Peptide Fragments / analysis
  • Phosphotransferases / isolation & purification*
  • Phosphotransferases / metabolism


  • Amino Acids
  • Detergents
  • Peptide Fragments
  • Phosphotransferases
  • Diacylglycerol Kinase
  • Cyanogen Bromide