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. 1976 Feb 1;153(2):463–468. doi: 10.1042/bj1530463

Purification and some properties of acetyl-coenzyme A carboxylase from rabbit mammary gland.

R Manning, R Dils, R J Mayer
PMCID: PMC1172594  PMID: 6012

Abstract

1. Acetyl-Coa carboxylase from lactating-rabbit mammary gland was purified to homogeneity by the criterion of polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. 2. Use of phosphate buffer throughout the purification gave low recovery of enzyme. Consequently, Tris buffers were used in the extraction and in selected stages of the purification procedure. 3. The purified enzyme had a specific activity of 5.15 +/- 0.3 mumol of bicarbonate incorporated/min per mg of protein (mean +/- S.E.M. of five preparations). This represents a purification of 257 +/- 16-fold and a yield of 4.3 +/- 0.13%. 4. The kinetic parameters of the purified enzyme were similar to those reported for the enzyme from other tissue sources. 5. The enzyme was assayed by a spectrophotometric assay and by a [14C]bicarbonate-fixation assay. Short incubation were used in the radio-chemical assay to avoid substantial loss of [14C]bicarbonate.

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Selected References

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

  1. Betts S. A., Mayer R. J. Purification and properties of 6-phosphogluconate dehydrogenase from rabbit mammary gland. Biochem J. 1975 Nov;151(2):263–270. doi: 10.1042/bj1510263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Carey E. M., Dils R. Fatty acid biosynthesis. V. Purification and characterisation of fatty acid synthetase from lactating-rabbit mammary gland. Biochim Biophys Acta. 1970 Sep 8;210(3):371–387. doi: 10.1016/0005-2760(70)90033-0. [DOI] [PubMed] [Google Scholar]
  3. Chase J. F. pH-dependence of carnitine acetyltransferase activity. Biochem J. 1967 Aug;104(2):503–509. doi: 10.1042/bj1040503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Gregolin C., Ryder E., Lane M. D. Liver acetyl coenzyme A carboxylase. I. Isolation and cat- alytic properties. J Biol Chem. 1968 Aug 25;243(16):4227–4235. [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. Guchhait R. B., Zwergel E. E., Lane M. D. Acetyl coenzyme A carboxylase. Subunit structure of the protomeric form of the avian liver enzyme. J Biol Chem. 1974 Aug 10;249(15):4776–4780. [PubMed] [Google Scholar]
  8. Hübscher G., West G. R., Brindley D. N. Studies on the fractionation of mucosal homogenates from the small intestine. Biochem J. 1965 Dec;97(3):629–642. doi: 10.1042/bj0970629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Inoue H., Lowenstein J. M. Acetyl coenzyme A carboxylase from rat liver. EC 6.4.1.2 acetyl-CoA: carbon dioxide ligase (ADP). Methods Enzymol. 1975;35:3–11. doi: 10.1016/0076-6879(75)35131-8. [DOI] [PubMed] [Google Scholar]
  10. Inoue H., Lowenstein J. M. Acetyl coenzyme A carboxylase from rat liver. Purification and demonstration of different subunits. J Biol Chem. 1972 Aug 10;247(15):4825–4832. [PubMed] [Google Scholar]
  11. 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]
  12. Lane M. D., Moss J., Polakis S. E. Acetyl coenzyme A carboxylase. Curr Top Cell Regul. 1974;8(0):139–195. [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. Majerus P. W., Jacobs R., Smith M. B., Morris H. P. The regulation of fatty acid biosynthesis in rat hepatomas. J Biol Chem. 1968 Jul 10;243(13):3588–3595. [PubMed] [Google Scholar]
  15. Majerus P. W., Kilburn E. Acetyl coenzyme A carboxylase. The roles of synthesis and degradation in regulation of enzyme levels in rat liver. J Biol Chem. 1969 Nov 25;244(22):6254–6262. [PubMed] [Google Scholar]
  16. Manning R., Brindley D. N. Tritium isotope effects in the measurement of the glycerol phosphate and dihydroxyacetone phosphate pathways of glycerolipid biosynthesis in rat liver. Biochem J. 1972 Dec;130(4):1003–1012. doi: 10.1042/bj1301003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Moss J., Yamagishi M., Kleinschmidt A. K., Lane M. D. Acetyl coenzyme A carboxylase. Purification and properties of the bovine adipose tissue enzyme. Biochemistry. 1972 Sep 26;11(20):3779–3786. doi: 10.1021/bi00770a017. [DOI] [PubMed] [Google Scholar]
  19. NATELSON S. Routine use of ultramicro methods in the clinical laboratory; estimation of sodium, potassium, chloride, protein, hematocrit value, sugar, urea and nonprotein nitrogen in fingertip blood; construction of ultramicro pipets; a practical microgasometer for estimation of carbon dioxide. Am J Clin Pathol. 1951 Dec;21(12):1153–1172. [PubMed] [Google Scholar]
  20. Nakanishi S., Numa S. Purification of rat liver acetyl coenzyme A carboxylase and immunochemical studies on its synthesis and degradation. Eur J Biochem. 1970 Sep;16(1):161–173. doi: 10.1111/j.1432-1033.1970.tb01068.x. [DOI] [PubMed] [Google Scholar]
  21. Numa S. Regulation of fatty-acid synthesis in higher animals. Ergeb Physiol. 1974;69(0):54–96. [PubMed] [Google Scholar]
  22. Smith S., Easter D. J., Dils R. Fatty acid biosynthesis. 3. Intracellular site of enzymes in lactating-rabbit mammary gland. Biochim Biophys Acta. 1966 Dec 7;125(3):445–455. [PubMed] [Google Scholar]
  23. WAITE M., WAKIL S. J. Studies on the mechanism of fatty acid synthesis. XII. Acetyl coenzyme A carboxylase. J Biol Chem. 1962 Sep;237:2750–2757. [PubMed] [Google Scholar]

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