Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1987 Apr;169(4):1469–1473. doi: 10.1128/jb.169.4.1469-1473.1987

Altered molecular form of acyl carrier protein associated with beta-ketoacyl-acyl carrier protein synthase II (fabF) mutants.

S Jackowski, C O Rock
PMCID: PMC211969  PMID: 3549687

Abstract

Acyl carrier protein (ACP) is a required cofactor for fatty acid synthesis in Escherichia coli. Mutants lacking beta-ketoacyl-ACP synthase II activity (fabF1 or fabF3) possessed a different molecular species of ACP (F-ACP) that was separated from the normal form of the protein by conformationally sensitive gel electrophoresis. Synthase I mutants contained the normal protein. Complementation of fabF1 mutants with an F' factor harboring the wild-type synthase II allele resulted in the appearance of normal ACP, whereas complementation with an F' possessing the fabF2 allele (a mutation that produces a synthase II enzyme with altered catalytic activity) resulted in the production of both forms of ACP. The structural difference between F-ACP and ACP persisted after the removal of the 4'-phosphopantetheine prosthetic group, and both forms of the protein had identical properties in an in vitro fatty acid synthase assay. Both ACP and F-ACP were purified to homogeneity, and their primary amino acid sequences were determined. The two ACP species were identical but differed from the sequence reported for E. coli E-15 ACP in that an Asn instead of an Asp was at position 24 and an Ile instead of a Val was at position 43. Therefore, F-ACP appears to be a modification of ACP that is detected when beta-ketoacyl-ACP synthase II activity is impaired.

Full text

PDF
1473

Images in this article

1470Image
on p.1470
1471Image
on p.1471
1471Image
on p.1471
1472Image
on p.1472

Selected References

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

  1. Bachmann B. J. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 1972 Dec;36(4):525–557. doi: 10.1128/br.36.4.525-557.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Booth B. R. Cell surface proteins of E. coli. Biochem Biophys Res Commun. 1980 Jun 30;94(4):1029–1036. doi: 10.1016/0006-291x(80)90522-7. [DOI] [PubMed] [Google Scholar]
  3. Cronan J. E., Jr, Weisberg L. J., Allen R. G. Regulation of membrane lipid synthesis in Escherichia coli. Accumulation of free fatty acids of abnormal length during inhibition of phospholipid synthesis. J Biol Chem. 1975 Aug 10;250(15):5835–5840. [PubMed] [Google Scholar]
  4. D'Agnolo G., Rosenfeld I. S., Vagelos P. R. Multiple forms of beta-ketoacyl-acyl carrier protein synthetase in Escherichia coli. J Biol Chem. 1975 Jul 25;250(14):5289–5294. [PubMed] [Google Scholar]
  5. Elovson J., Vagelos P. R. Acyl carrier protein. X. Acyl carrier protein synthetase. J Biol Chem. 1968 Jul 10;243(13):3603–3611. [PubMed] [Google Scholar]
  6. Garwin J. L., Klages A. L., Cronan J. E., Jr Beta-ketoacyl-acyl carrier protein synthase II of Escherichia coli. Evidence for function in the thermal regulation of fatty acid synthesis. J Biol Chem. 1980 Apr 25;255(8):3263–3265. [PubMed] [Google Scholar]
  7. Garwin J. L., Klages A. L., Cronan J. E., Jr Structural, enzymatic, and genetic studies of beta-ketoacyl-acyl carrier protein synthases I and II of Escherichia coli. J Biol Chem. 1980 Dec 25;255(24):11949–11956. [PubMed] [Google Scholar]
  8. Gelmann E. P., Cronan J. E., Jr Mutant of Escherichia coli deficient in the synthesis of cis-vaccenic acid. J Bacteriol. 1972 Oct;112(1):381–387. doi: 10.1128/jb.112.1.381-387.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jackowski S., Rock C. O. Consequences of reduced intracellular coenzyme A content in Escherichia coli. J Bacteriol. 1986 Jun;166(3):866–871. doi: 10.1128/jb.166.3.866-871.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. 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]
  12. Maloy S. R., Nunn W. D. Selection for loss of tetracycline resistance by Escherichia coli. J Bacteriol. 1981 Feb;145(2):1110–1111. doi: 10.1128/jb.145.2.1110-1111.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mayo K. H., Prestegard J. H. Acyl carrier protein from Escherichia coli. Structural characterization of short-chain acylated acyl carrier proteins by NMR. Biochemistry. 1985 Dec 17;24(26):7834–7838. doi: 10.1021/bi00347a049. [DOI] [PubMed] [Google Scholar]
  14. Mayo K. H., Tyrell P. M., Prestegard J. H. Acyl carrier protein from Escherichia coli I. Aspects of the solution structure as evidenced by proton nuclear Overhauser experiments at 500 MHz. Biochemistry. 1983 Sep 13;22(19):4485–4493. doi: 10.1021/bi00288a021. [DOI] [PubMed] [Google Scholar]
  15. McRee D. E., Richardson J. S., Richardson D. C. Preliminary X-ray diffraction studies of acyl carrier protein from Escherichia coli. J Mol Biol. 1985 Apr 5;182(3):467–468. doi: 10.1016/0022-2836(85)90205-0. [DOI] [PubMed] [Google Scholar]
  16. Okuyama H., Saitoh M., Hiramatsu R. Fatty acid synthetase system in the regulation of membrane lipid synthesis in Escherichia coli after shifts in temperature. J Biol Chem. 1982 May 10;257(9):4812–4817. [PubMed] [Google Scholar]
  17. Prescott D. J., Vagelos P. R. Acyl carrier protein. Adv Enzymol Relat Areas Mol Biol. 1972;36:269–311. doi: 10.1002/9780470122815.ch8. [DOI] [PubMed] [Google Scholar]
  18. Rock C. O., Cronan J. E., Jr, Armitage I. M. Molecular properties of acyl carrier protein derivatives. J Biol Chem. 1981 Mar 25;256(6):2669–2674. [PubMed] [Google Scholar]
  19. Rock C. O., Cronan J. E., Jr Re-evaluation of the solution structure of acyl carrier protein. J Biol Chem. 1979 Oct 10;254(19):9778–9785. [PubMed] [Google Scholar]
  20. Rock C. O. Environment of the aromatic chromophores of acyl carrier protein. Arch Biochem Biophys. 1983 Aug;225(1):122–129. doi: 10.1016/0003-9861(83)90014-0. [DOI] [PubMed] [Google Scholar]
  21. Rock C. O. Mixed disulfides of acyl carrier protein and coenzyme A with specific soluble proteins in Escherichia coli. J Bacteriol. 1982 Dec;152(3):1298–1300. doi: 10.1128/jb.152.3.1298-1300.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rosenfeld I. S., D'Agnolo G., Vagelos P. R. Synthesis of unsaturated fatty acids and the lesion in fab B mutants. J Biol Chem. 1973 Apr 10;248(7):2452–2460. [PubMed] [Google Scholar]
  23. Ulrich A. K., de Mendoza D., Garwin J. L., Cronan J. E., Jr Genetic and biochemical analyses of Escherichia coli mutants altered in the temperature-dependent regulation of membrane lipid composition. J Bacteriol. 1983 Apr;154(1):221–230. doi: 10.1128/jb.154.1.221-230.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
  25. 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]
  26. Vanaman T. C., Wakil S. J., Hill R. L. The complete amino acid sequence of the acyl carrier protein of Escherichia coli. J Biol Chem. 1968 Dec 25;243(24):6420–6431. [PubMed] [Google Scholar]
  27. Weeks G., Wakil S. J. Studies on the mechanism of fatty acid synthesis. 18. Preparation and general properties of the enoyl acyl carrier protein reductases from Escherichia coli. J Biol Chem. 1968 Mar 25;243(6):1180–1189. [PubMed] [Google Scholar]
  28. de Mendoza D., Garwin J. L., Cronan J. E., Jr Overproduction of cis-vaccenic acid and altered temperature control of fatty acid synthesis in a mutant of Escherichia coli. J Bacteriol. 1982 Sep;151(3):1608–1611. doi: 10.1128/jb.151.3.1608-1611.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES