Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1983 Oct 1;215(1):153–157. doi: 10.1042/bj2150153

A bifunctional enzyme complex in coenzyme A biosynthesis: purification of pantetheine phosphate adenylyltransferase and dephospho-CoA kinase.

D M Worrall, P K Tubbs
PMCID: PMC1152375  PMID: 6312972

Abstract

Pantetheine phosphate adenylyltransferase (EC 2.7.7.3) and dephospho-CoA kinase (EC 2.7.1.24) were purified to near homogeneity from pig liver. The purification steps included the use of Sepharose-linked triazine dyes and affinity elution by CoA. Both activities co-purified at every stage of the 18 000-fold purification. An Mr of 115 000 was obtained by gel filtration on Sephadex G-150, and the final preparation yielded one major band on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, with a subunit Mr of 57 000. It is concluded that pantetheine phosphate adenylyltransferase and dephospho-CoA kinase exist as a bifunctional dimeric protein, which could be designated CoA synthetase.

Full text

PDF
154

Images in this article

156Fig. 3.
on p.156

Selected References

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

  1. Baird J. K., Sherwood R. F., Carr R. J., Atkinson A. Enzyme purification by substrate elution chromatography from procion dye-polysaccharide matrices. FEBS Lett. 1976 Nov;70(1):61–66. doi: 10.1016/0014-5793(76)80726-0. [DOI] [PubMed] [Google Scholar]
  2. HOAGLAND M. B., NOVELLI G. D. Biosynthesis of coenzyme A from phospho-pantetheine and of pantetheine from pantothenate. J Biol Chem. 1954 Apr;207(2):767–773. [PubMed] [Google Scholar]
  3. Heinonen J. K., Lahti R. J. A new and convenient colorimetric determination of inorganic orthophosphate and its application to the assay of inorganic pyrophosphatase. Anal Biochem. 1981 May 15;113(2):313–317. doi: 10.1016/0003-2697(81)90082-8. [DOI] [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. Peterson G. L. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem. 1977 Dec;83(2):346–356. doi: 10.1016/0003-2697(77)90043-4. [DOI] [PubMed] [Google Scholar]
  6. Skrede S., Halvorsen O. Mitochondrial biosynthesis of coenzyme A. Biochem Biophys Res Commun. 1979 Dec 28;91(4):1536–1542. doi: 10.1016/0006-291x(79)91239-7. [DOI] [PubMed] [Google Scholar]
  7. Skrede S., Halvorsen O. Mitochondrial pantetheinephosphate adenylyltransferase and dephospho-CoA kinase. Eur J Biochem. 1983 Mar 1;131(1):57–63. doi: 10.1111/j.1432-1033.1983.tb07231.x. [DOI] [PubMed] [Google Scholar]
  8. Suzuki T., Abiko Y., Shimizu M. Investigations on pantothenic acid and its related compounds. XII. Biochemical studies (7). Dephospho-coA pyrophosphorylase and dephospho-coA kinase as a possible bifunctional enzyme complex. J Biochem. 1967 Dec;62(6):642–649. doi: 10.1093/oxfordjournals.jbchem.a128719. [DOI] [PubMed] [Google Scholar]
  9. WANG T. P., SHUSTER L., KAPLAN N. O. The monoester phosphate grouping of coenzyme A. J Biol Chem. 1954 Jan;206(1):299–309. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES