The Escherichia coli pgpB gene encodes for a diacylglycerol pyrophosphate phosphatase activity

Deirdre A. Dillon, Wen I. Wut, Bettina Riedel, Josef B. Wissing, William Dowhan, George Carman

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Abstract

We provided genetic and biochemical evidence that supported the conclusion that the product of pgpB gene of Escherichia coli exhibited diacylglycerol pyrophosphate (DGPP) phosphatase activity. DGPP phosphatase activity was absent in pgpB mutant cells and was expressed at high levels in cells carrying the wild-type pgpB gene on a runaway replication plasmid. The pgpB mutant has been primarily characterized by a defect in phosphatidate (PA) phosphatase activity and also exhibits defects in lyso-PA phosphatase and phosphatidylglycerophosphate phosphatase activities. The defective PA phosphatase in the pgpB mutant was shown to be a Mg2+-independent PA phosphatase activity of the DGPP phosphatase enzyme. We characterized DGPP phosphatase activity in membranes from cells overproducing the pgpB gene product. DGPP phosphatase catalyzed the dephosphorylation of the β phosphate of DGPP to form PA followed by the dephosphorylation of PA to form diacylglycerol. The specificity constant (V(max)/K(m)) for DGPP was 9.3-fold greater than that for PA. The pH optimum for the DGPP phosphatase reaction was 6.5. Activity was independent of a divalent cation requirement, was potently inhibited by Mn2+ ions, and was insensitive to inhibition by N- ethylmaleimide. Pure DGPP phosphatase from Saccharomyces cerevisiae was shown to be similar to the E. coli DGPP phosphatase in its ability to utilize lyso- PA and phosphatidylglycerophosphate as substrates in vitro.

Original languageEnglish (US)
Pages (from-to)30548-30553
Number of pages6
JournalJournal of Biological Chemistry
Volume271
Issue number48
DOIs
StatePublished - Dec 14 1996

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Escherichia coli
Genes
Phosphatidate Phosphatase
Homeless Youth
Defects
Ethylmaleimide
diacylglycerol pyrophosphate phosphatase
Diglycerides
Divalent Cations
Phosphoric Monoester Hydrolases
Yeast
Saccharomyces cerevisiae
Molecular Biology
Plasmids
Phosphates
Cell Membrane
Ions
Membranes
Substrates
Enzymes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Dillon, Deirdre A. ; Wut, Wen I. ; Riedel, Bettina ; Wissing, Josef B. ; Dowhan, William ; Carman, George. / The Escherichia coli pgpB gene encodes for a diacylglycerol pyrophosphate phosphatase activity. In: Journal of Biological Chemistry. 1996 ; Vol. 271, No. 48. pp. 30548-30553.
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The Escherichia coli pgpB gene encodes for a diacylglycerol pyrophosphate phosphatase activity. / Dillon, Deirdre A.; Wut, Wen I.; Riedel, Bettina; Wissing, Josef B.; Dowhan, William; Carman, George.

In: Journal of Biological Chemistry, Vol. 271, No. 48, 14.12.1996, p. 30548-30553.

Research output: Contribution to journalArticle

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T1 - The Escherichia coli pgpB gene encodes for a diacylglycerol pyrophosphate phosphatase activity

AU - Dillon, Deirdre A.

AU - Wut, Wen I.

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