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. 1990 Sep;172(9):5445–5449. doi: 10.1128/jb.172.9.5445-5449.1990

Isolation and properties of acyl carrier protein phosphodiesterase of Escherichia coli.

A S Fischl 1, E P Kennedy 1
PMCID: PMC213211  PMID: 2168383

Abstract

The acyl carrier protein (ACP) phosphodiesterase of Escherichia coli catalyzes the hydrolytic cleavage of the 4'-phosphopantetheine residue from ACP, with the generation of apo-ACP (P. R. Vagelos and A. R. Larrabee, J. Biol. Chem. 242:1776-1781, 1967). Although it has been postulated to play a role in the regulation of fatty acid synthesis, presently available evidence makes this unlikely, and its physiological function requires further investigation. We have now purified the enzyme from E. coli more than 3,000-fold and have identified it as a protein of Mr 25,000, as judged from its migration during electrophoresis in gels containing sodium dodecyl sulfate. The enzyme has remarkable thermostability, being protected against irreversible inactivation at 90 degrees C by the presence of sodium dodecyl sulfate. A partial sequence of the amino terminus of the enzyme is as follows: H2N-Ser-Lys-Val-Leu-Val-Leu-Lys-Ser-?-Ile-Leu-Ala-Gly-Tyr-Ser-. Other properties of the enzyme are also described.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson M. S., Raetz C. R. Biosynthesis of lipid A precursors in Escherichia coli. A cytoplasmic acyltransferase that converts UDP-N-acetylglucosamine to UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine. J Biol Chem. 1987 Apr 15;262(11):5159–5169. [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  3. Cooper C. L., Hsu L., Jackowski S., Rock C. O. 2-Acylglycerolphosphoethanolamine acyltransferase/acyl-acyl carrier protein synthetase is a membrane-associated acyl carrier protein binding protein. J Biol Chem. 1989 May 5;264(13):7384–7389. [PubMed] [Google Scholar]
  4. Hathcock K. S., Gurish M. F., Nisonoff A., Conger J. D., Hodes R. J. Influence of helper T cells on the expression of a murine intrastrain crossreactive idiotype. Proc Natl Acad Sci U S A. 1986 Jan;83(1):155–159. doi: 10.1073/pnas.83.1.155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Jackowski S., Rock C. O. Ratio of active to inactive forms of acyl carrier protein in Escherichia coli. J Biol Chem. 1983 Dec 25;258(24):15186–15191. [PubMed] [Google Scholar]
  6. Jackowski S., Rock C. O. Regulation of coenzyme A biosynthesis. J Bacteriol. 1981 Dec;148(3):926–932. doi: 10.1128/jb.148.3.926-932.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Jackowski S., Rock C. O. Turnover of the 4'-phosphopantetheine prosthetic group of acyl carrier protein. J Biol Chem. 1984 Feb 10;259(3):1891–1895. [PubMed] [Google Scholar]
  8. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  9. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  10. March S. C., Parikh I., Cuatrecasas P. A simplified method for cyanogen bromide activation of agarose for affinity chromatography. Anal Biochem. 1974 Jul;60(1):149–152. doi: 10.1016/0003-2697(74)90139-0. [DOI] [PubMed] [Google Scholar]
  11. Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem. 1987 Jul 25;262(21):10035–10038. [PubMed] [Google Scholar]
  12. Powell G. L., Elovson J., Vagelos P. R. Acyl carrier protein. XII. Synthesis and turnover of the prosthetic group of acyl carrier protein in vivo. J Biol Chem. 1969 Oct 25;244(20):5616–5624. [PubMed] [Google Scholar]
  13. Ryan L. D., Vestling C. S. Rapid purification of lactate dehydrogenase from rat liver and hepatoma: a new approach. Arch Biochem Biophys. 1974 Jan;160(1):279–284. doi: 10.1016/s0003-9861(74)80035-4. [DOI] [PubMed] [Google Scholar]
  14. Therisod H., Kennedy E. P. The function of acyl carrier protein in the synthesis of membrane-derived oligosaccharides does not require its phosphopantetheine prosthetic group. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8235–8238. doi: 10.1073/pnas.84.23.8235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Vagelos P. R., Larrabes A. R. Acyl carrier protein. IX. Acyl carrier protein hydrolase. J Biol Chem. 1967 Apr 25;242(8):1776–1781. [PubMed] [Google Scholar]

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