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. 1984 Aug;159(2):720–724. doi: 10.1128/jb.159.2.720-724.1984

In vivo and in vitro acylation of polypeptides in Vibrio harveyi: identification of proteins involved in aldehyde production for bioluminescence.

L A Wall, D M Byers, E A Meighen
PMCID: PMC215704  PMID: 6746576

Abstract

Incubation of soluble extracts from Vibrio harveyi with [3H]tetradecanoic acid (+ ATP) resulted in the acylation of several polypeptides, including proteins with molecular masses near 20 kilodaltons (kDa), and at least five polypeptides in the 30- to 60-kDa range. However, in growing cells pulse-labeled in vivo with [3H]tetradecanoic acid, only three of these polypeptides, with apparent molecular masses of 54, 42, and 32 kDa, were specifically labeled. When extracts were acylated with [3H] tetradecanoyl coenzyme A, on the other hand, only the 32-kDa polypeptide was labeled. When luciferase-containing dark mutants of V. harveyi were investigated, acylated 32-kDa polypeptide was not detected in a fatty acid-stimulated mutant, whereas the 42-kDa polypeptide appeared to be lacking in a mutant defective in aldehyde synthesis. Acylation of both of these polypeptides also increased specifically during induction of bioluminescence in V. harveyi. These results suggest that the role of the 32-kDa polypeptide is to supply free fatty acids, whereas the 42-kDa protein may be responsible for activation of fatty acids for their subsequent reduction to form the aldehyde substrates of the bioluminescent reaction.

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Selected References

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

  1. 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: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  2. Dunn D. K., Michaliszyn G. A., Bogacki I. G., Meighen E. A. Conversion of aldehyde to acid in the bacterial bioluminescent reaction. Biochemistry. 1973 Nov 20;12(24):4911–4918. doi: 10.1021/bi00748a016. [DOI] [PubMed] [Google Scholar]
  3. Gunsalus-Miguel A., Meighen E. A., Nicoli M. Z., Nealson K. H., Hastings J. W. Purification and properties of bacterial luciferases. J Biol Chem. 1972 Jan 25;247(2):398–404. [PubMed] [Google Scholar]
  4. 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]
  5. Meighen E. A., Bartlet I. Complementation of subunits from different bacterial luciferases. Evidence for the role of the beta subunit in the bioluminescent mechanism. J Biol Chem. 1980 Dec 10;255(23):11181–11187. [PubMed] [Google Scholar]
  6. Meighen E. A. Biosynthesis of aliphatic aldehydes for the bacterial bioluminescent reaction: stimulation by ATP and NADPH. Biochem Biophys Res Commun. 1979 Apr 27;87(4):1080–1086. doi: 10.1016/s0006-291x(79)80018-2. [DOI] [PubMed] [Google Scholar]
  7. Michaliszyn G. A., Meighen E. A. Induced polypeptide synthesis during the development of bacterial bioluminescence. J Biol Chem. 1976 May 10;251(9):2541–2549. [PubMed] [Google Scholar]
  8. Nealson K. H., Platt T., Hastings J. W. Cellular control of the synthesis and activity of the bacterial luminescent system. J Bacteriol. 1970 Oct;104(1):313–322. doi: 10.1128/jb.104.1.313-322.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ray T. K., Cronan J. E., Jr Activation of long chain fatty acids with acyl carrier protein: demonstration of a new enzyme, acyl-acyl carrier protein synthetase, in Escherichia coli. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4374–4378. doi: 10.1073/pnas.73.12.4374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Riendeau D., Meighen E. Evidence for a fatty acid reductase catalyzing the synthesis of aldehydes for the bacterial bioluminescent reaction. Resolution from luciferase and dependence on fatty acids. J Biol Chem. 1979 Aug 25;254(16):7488–7490. [PubMed] [Google Scholar]
  11. 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]
  12. Rodriguez A., Riendeau D., Meighen E. Purification of the acyl coenzyme A reductase component from a complex responsible for the reduction of fatty acids in bioluminescent bacteria. Properties and acyltransferase activity. J Biol Chem. 1983 Apr 25;258(8):5233–5237. [PubMed] [Google Scholar]
  13. Silbert D. F., Ruch F., Vagelos P. R. Fatty acid replacements in a fatty acid auxotroph of Escherichia coli. J Bacteriol. 1968 May;95(5):1658–1665. doi: 10.1128/jb.95.5.1658-1665.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ulitzur S., Hastings J. W. Myristic acid stimulation of bacterial bioluminescence in "aldehyde" mutants. Proc Natl Acad Sci U S A. 1978 Jan;75(1):266–269. doi: 10.1073/pnas.75.1.266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Wall L., Rodriquez A., Meighen E. Differential acylation in vitro with tetradecanoyl coenzyme A and tetradecanoic acid (+ATP) of three polypeptides shown to have induced synthesis in Photobacterium phosphoreum. J Biol Chem. 1984 Feb 10;259(3):1409–1414. [PubMed] [Google Scholar]

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