Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1970 Nov;67(3):1353–1360. doi: 10.1073/pnas.67.3.1353

Enzymatic Carboxylation of Biotin: Molecular and Catalytic Properties of a Component Enzyme of Acetyl CoA Carboxylase*

Peter Dimroth 1,2, Ras B Guchhait 1,2, Erwin Stoll 1,2, M Daniel Lane 1,2
PMCID: PMC283359  PMID: 4922289

Abstract

The biotin carboxylase component of acetyl CoA carboxylase has been purified approximately 2000 times from Escherichia coli. This protein, which catalyzes the carboxylation of free d-biotin, is free of the biotin-containing carboxyl carrier protein, is homogeneous by polyacrylamide gel electrophoresis and analytical ultracentrifugation, and has been crystallized. Biotin carboxylase, with a molecular weight of approximately 100,000, is composed of two 50,000-dalton subunits. The catalytic capacity of biotin carboxylase is markedly enhanced by ethanol (11 times at 15% v/v), and certain other organic solvents; this may mimic an effector-mediated response. The kinetic effect is exclusively on the maximal velocity of the reaction. Activation by ethanol is reversible and not accompanied by aggregation or disaggregation of the enzyme.

Full text

PDF
1353

Images in this article

1356Image
on p.1356

Selected References

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

  1. Alberts A. W., Nervi A. M., Vagelos P. R. Acetyl CoA carboxylase, II. Deomonstration of biotin-protein and biotin carboxylase subunits. Proc Natl Acad Sci U S A. 1969 Aug;63(4):1319–1326. doi: 10.1073/pnas.63.4.1319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alberts A. W., Vagelos P. R. Acetyl CoA carboxylase. I. Requirement for two protein fractions. Proc Natl Acad Sci U S A. 1968 Feb;59(2):561–568. doi: 10.1073/pnas.59.2.561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cuatrecasas P., Wilchek M. Single-step purification of avidine from egg white by affinity chromatography on biocytin-Sepharose columns. Biochem Biophys Res Commun. 1968 Oct 24;33(2):235–239. doi: 10.1016/0006-291x(68)90774-2. [DOI] [PubMed] [Google Scholar]
  4. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  5. Gregolin C., Ryder E., Kleinschmidt A. K., Warner R. C., Lane M. D. Molecular characteristics of liver acetyl CoA carboxylase. Proc Natl Acad Sci U S A. 1966 Jul;56(1):148–155. doi: 10.1073/pnas.56.1.148. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gregolin C., Ryder E., Warner R. C., Kleinschmidt A. K., Chang H. C., Lane M. D. Liver acetyl coenzyme A carboxylase. II. Further molecular characterization. J Biol Chem. 1968 Aug 25;243(16):4236–4245. [PubMed] [Google Scholar]
  7. Gregolin C., Ryder E., Warner R. C., Kleinschmidt A. K., Lane M. D. Liver acetyl coa carboxylase: the dissociation-reassociation process and its relation to catalytic activity. Proc Natl Acad Sci U S A. 1966 Dec;56(6):1751–1758. doi: 10.1073/pnas.56.6.1751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jovin T. M., Englund P. T., Bertsch L. L. Enzymatic synthesis of deoxyribonucleic acid. XXVI. Physical and chemical studies of a homogeneous deoxyribonucleic acid polymerase. J Biol Chem. 1969 Jun 10;244(11):2996–3008. [PubMed] [Google Scholar]
  9. KAZIRO Y., OCHOA S. THE METABOLISM OF PROPIONIC ACID. Adv Enzymol Relat Areas Mol Biol. 1964;26:283–378. doi: 10.1002/9780470122716.ch7. [DOI] [PubMed] [Google Scholar]
  10. KNAPPE J., RINGELMANN E., LYNEN F. [On the biochemical function of biotin. III. The chemical structure of enzymatically formed carboxy-biotins]. Biochem Z. 1961;335:168–176. [PubMed] [Google Scholar]
  11. Knivett V. A., Cullen J. Fatty acid synthesis in Escherichia coli. Biochem J. 1967 May;103(2):299–306. doi: 10.1042/bj1030299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. LYNEN F., KNAPPE J., LORCH E., JUETTING G., RINGELMANN E., LACHANCE J. P. [On the biochemical function of biotin. II. Purification and mode of action of beta-methyl-crotonyl-carboxylase]. Biochem Z. 1961;335:123–167. [PubMed] [Google Scholar]
  13. MATSUHASHI M., MATSUHASHI S., NUMA S., LYNEN F. ZUR BIOSYNTHESE DER FETTSAEUREN. IV. ACETYL-COA CARBOXYLASE AUS HEFE. Biochem Z. 1964 Aug 11;340:243–262. [PubMed] [Google Scholar]
  14. Miller A. L., Levy H. R. Rat mammary acetyl coenzyme A carboxylase. I. Isolation and characterization. J Biol Chem. 1969 May 10;244(9):2334–2342. [PubMed] [Google Scholar]
  15. Numa S., Ringelmann E., Riedel B. Further evidence for polymeric structure of liver acetyl CoA carboxylase. Biochem Biophys Res Commun. 1966 Sep 8;24(5):750–757. doi: 10.1016/0006-291x(66)90389-5. [DOI] [PubMed] [Google Scholar]
  16. Roark D. E., Yphantis D. A. Studies of self-associating systems by equilibrium ultracentrifugation. Ann N Y Acad Sci. 1969 Nov 7;164(1):245–278. doi: 10.1111/j.1749-6632.1969.tb14043.x. [DOI] [PubMed] [Google Scholar]
  17. Rutner A. C. Estimation of the molecular weight of ribulose diphosphate carboxylase sub-units. Biochem Biophys Res Commun. 1970 Jun 5;39(5):923–929. doi: 10.1016/0006-291x(70)90412-2. [DOI] [PubMed] [Google Scholar]
  18. Sanwal B. D., Maeba P., Cook R. A. Interaction of macroions and dioxane with the allosteric phosphoenolpyruvate carboxylase. J Biol Chem. 1966 Nov 25;241(22):5177–5182. [PubMed] [Google Scholar]
  19. Siegel L. M., Monty K. J. Determination of molecular weights and frictional ratios of proteins in impure systems by use of gel filtration and density gradient centrifugation. Application to crude preparations of sulfite and hydroxylamine reductases. Biochim Biophys Acta. 1966 Feb 7;112(2):346–362. doi: 10.1016/0926-6585(66)90333-5. [DOI] [PubMed] [Google Scholar]
  20. Stoll E., Ryder E., Edwards J. B., Lane M. D. Liver acetyl coenzyme A carboxylase: activation of model partial reactions by tricarboxylic acids. Proc Natl Acad Sci U S A. 1968 Jul;60(3):986–991. doi: 10.1073/pnas.60.3.986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. WAKIL S. J., TITCHENER E. B., GIBSON D. M. Evidence for the participation of biotin in the enzymic synthesis of fatty acids. Biochim Biophys Acta. 1958 Jul;29(1):225–226. doi: 10.1016/0006-3002(58)90177-x. [DOI] [PubMed] [Google Scholar]
  22. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES