Abstract
Acetyl-CoA carboxylase from the diatom Cyclotella cryptica has been purified to near homogeneity by the use of ammonium sulfate fractionation, gel filtration chromatography, and affinity chromatography with monomeric avidin-agarose. The specific activity of the final preparation was as high as 14.6 micromoles malonyl-CoA formed per milligram protein per minute, indicating a 600-fold purification. Native acetyl-CoA carboxylase has a molecular weight of approximately 740 kilodaltons and appears to be composed of four identical biotin-containing subunits. The enzyme has maximal activity at pH 8.2, but enzyme stability is greater at pH 6.5. Km values for MgATP, acetyl-CoA, and HCO3- were determined to be 65, 233, and 750 micromolar, respectively. The purified enzyme is strongly inhibited by palmitoyl-CoA, and is inhibited to a lesser extent by malonyl-CoA, ADP, and phosphate. Pyruvate stimulates enzymatic activity to a slight extent. Acetyl-CoA carboxylase from Cyclotella cryptica is not inhibited by cyclohexanedione or aryloxyphenoxypropionic acid herbicides as strongly as monocot acetyl-CoA carboxylases; 50% and 0% inhibition was observed in the presence of 23 micromolar clethodim and 100 micromolar haloxyfop, respectively.
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- Burton J. D., Gronwald J. W., Somers D. A., Connelly J. A., Gengenbach B. G., Wyse D. L. Inhibition of plant acetyl-coenzyme A carboxylase by the herbicides sethoxydim and haloxyfop. Biochem Biophys Res Commun. 1987 Nov 13;148(3):1039–1044. doi: 10.1016/s0006-291x(87)80236-x. [DOI] [PubMed] [Google Scholar]
- Eastwell K. C., Stumpf P. K. Regulation of Plant Acetyl-CoA Carboxylase by Adenylate Nucleotides. Plant Physiol. 1983 May;72(1):50–55. doi: 10.1104/pp.72.1.50. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Egin-Bühler B., Ebel J. Improved purification and further characterization of acetyl-CoA carboxylase from cultured cells of parsley (Petroselinum hortense). Eur J Biochem. 1983 Jun 15;133(2):335–339. doi: 10.1111/j.1432-1033.1983.tb07467.x. [DOI] [PubMed] [Google Scholar]
- Egin-Bühler B., Loyal R., Ebel J. Comparison of acetyl-CoA carboxylases from parsley cell cultures and wheat germ. Arch Biochem Biophys. 1980 Aug;203(1):90–100. doi: 10.1016/0003-9861(80)90156-3. [DOI] [PubMed] [Google Scholar]
- Finlayson S. A., Dennis D. T. Acetyl-coenzyme A carboxylase from the developing endosperm of Ricinus communis. I. Isolation and characterization. Arch Biochem Biophys. 1983 Sep;225(2):576–585. doi: 10.1016/0003-9861(83)90069-3. [DOI] [PubMed] [Google Scholar]
- Heinstein P. F., Stumpf P. K. Fat metabolism in higher plants. 38. Properties of wheat germ acetyl coenzyme A carboxylase. J Biol Chem. 1969 Oct 10;244(19):5374–5381. [PubMed] [Google Scholar]
- Hellyer A., Bambridge H. E., Slabas A. R. Plant acetyl-CoA carboxylase. Biochem Soc Trans. 1986 Jun;14(3):565–568. doi: 10.1042/bst0140565. [DOI] [PubMed] [Google Scholar]
- Henrikson K. P., Allen S. H., Maloy W. L. An avidin monomer affinity column for the purification of biotin-containing enzymes. Anal Biochem. 1979 Apr 15;94(2):366–370. doi: 10.1016/0003-2697(79)90374-9. [DOI] [PubMed] [Google Scholar]
- 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]
- Mohan S. B., Kekwick R. G. Acetyl-coenzyme A carboxylase from avocado (Persea americana) plastids and spinach (Spinacia oleracea) chloroplasts. Biochem J. 1980 Jun 1;187(3):667–676. doi: 10.1042/bj1870667. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nielsen N. C., Adee A., Stumpf P. K. Fat metabolism in higher plants. Further characterization of wheat germ acetyl coenzyme A carboxylase. Arch Biochem Biophys. 1979 Feb;192(2):446–456. doi: 10.1016/0003-9861(79)90114-0. [DOI] [PubMed] [Google Scholar]
- Nikolau B. J., Hawke J. C. Purification and characterization of maize leaf acetyl-coenzyme A carboxylase. Arch Biochem Biophys. 1984 Jan;228(1):86–96. doi: 10.1016/0003-9861(84)90049-3. [DOI] [PubMed] [Google Scholar]
- Nikolau B. J., Hawke J. C., Slack C. R. Acetyl-coenzyme A carboxylase in maize leaves. Arch Biochem Biophys. 1981 Oct 15;211(2):605–612. doi: 10.1016/0003-9861(81)90495-1. [DOI] [PubMed] [Google Scholar]
- Rendina A. R., Felts J. M., Beaudoin J. D., Craig-Kennard A. C., Look L. L., Paraskos S. L., Hagenah J. A. Kinetic characterization, stereoselectivity, and species selectivity of the inhibition of plant acetyl-CoA carboxylase by the aryloxyphenoxypropionic acid grass herbicides. Arch Biochem Biophys. 1988 Aug 15;265(1):219–225. doi: 10.1016/0003-9861(88)90387-6. [DOI] [PubMed] [Google Scholar]
- Rendina A. R., Felts J. M. Cyclohexanedione Herbicides Are Selective and Potent Inhibitors of Acetyl-CoA Carboxylase from Grasses. Plant Physiol. 1988 Apr;86(4):983–986. doi: 10.1104/pp.86.4.983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Roessler P. G. Changes in the activities of various lipid and carbohydrate biosynthetic enzymes in the diatom Cyclotella cryptica in response to silicon deficiency. Arch Biochem Biophys. 1988 Dec;267(2):521–528. doi: 10.1016/0003-9861(88)90059-8. [DOI] [PubMed] [Google Scholar]
- Schafer-Nielsen C., Svendsen P. J., Rose C. Separation of macromolecules in isotachophoresis systems involving single or multiple counterions. J Biochem Biophys Methods. 1980 Aug;3(2):97–128. doi: 10.1016/0165-022x(80)90032-9. [DOI] [PubMed] [Google Scholar]
- Secor J., Cséke C. Inhibition of Acetyl-CoA Carboxylase Activity by Haloxyfop and Tralkoxydim. Plant Physiol. 1988 Jan;86(1):10–12. doi: 10.1104/pp.86.1.10. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sumper M., Riepertinger C. Structural relationship of biotin-containing enzymes. Acetyl-CoA carboxylase and pyruvate carboxylase from yeast. Eur J Biochem. 1972 Sep 18;29(2):237–248. doi: 10.1111/j.1432-1033.1972.tb01980.x. [DOI] [PubMed] [Google Scholar]
- Thomson L. W., Zalik S. Acetyl coenzyme a carboxylase activity in developing seedlings and chloroplasts of barley and its virescens mutant. Plant Physiol. 1981 Apr;67(4):655–661. doi: 10.1104/pp.67.4.655. [DOI] [PMC free article] [PubMed] [Google Scholar]