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. 1975 Apr;72(4):1617–1621. doi: 10.1073/pnas.72.4.1617

Beta-cyanoalanine synthase: purification and characterization.

T N Akopyan, A E Braunstein, E V Goryachenkova
PMCID: PMC432590  PMID: 1055433

Abstract

Beta-cyano-L-alanine synthase [L-cysteine hydrogen-sulfide-lyase (adding HCN), EC 4.4.1.9] was purified about 4000-fold from blue lupine seedlings. The enzyme was homoegeneous on gel electrophoresis and free of contamination by other pyridoxal-P-dependent lyases. The enzyme has a molecular weight of 52,000 and contains 1 mole of pyridoxal-P per mole of protein; its isoelectric point is situated at pH 4.7. Its absorption spectrum has two maxima, at 280 and 410 nm. L-Cysteine is the natural primary (amino acid) substrate; beta-chloro- and beta-thiocyano can serve (with considerably lower affinity) instead of cyanide as cosubstrates for cyanoalanine synthase. The synthase is refractory to DL-cycloserine and D-penicillamine, potent inhibitors of many pyridoxal-P-dependent enzymes. Cyanoalanine synthase catalyzes slow isotopic alpha-H exchange in cysteine and in end-product amino acids; the rates of alpha-H exchange in nonreacted (excess) cysteine are markedly increased in the presence of an adequate cosubstrate; no exchange is observed of H atoms in beta-position.

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Selected References

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

  1. Andrews P. The gel-filtration behaviour of proteins related to their molecular weights over a wide range. Biochem J. 1965 Sep;96(3):595–606. doi: 10.1042/bj0960595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BODE F. Eine Vereinfachung und Verbesserung der Methode zur quantitativen Bestimmung von Aminosäuren und Peptiden mittels des Ninhydrin-Kupferkomplexes. Biochem Z. 1955;326(6):433–435. [PubMed] [Google Scholar]
  3. Braunstein A. E., Goryachenkova E. V., Lac N. D. Reactions catalysed by serine sulfhydrase from chicken liver. Biochim Biophys Acta. 1969 Feb 11;171(2):366–368. doi: 10.1016/0005-2744(69)90173-9. [DOI] [PubMed] [Google Scholar]
  4. Braunstein A. E., Goryachenkova E. V., Tolosa E. A., Willhardt I. H., Yefremova L. L. Specificity and some other properties of liver serine sulphhydrase: evidence for its identity with cystathionine -synthase. Biochim Biophys Acta. 1971 Jul 21;242(1):247–260. doi: 10.1016/0005-2744(71)90105-7. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Dunnill P. M., Fowden L. Enzymatic formation of beta-cyanoalanine from cyanide by Escherichia coli extracts. Nature. 1965 Dec 18;208(5016):1206–1207. doi: 10.1038/2081206a0. [DOI] [PubMed] [Google Scholar]
  7. Floss H. G., Hadwiger L., Conn E. E. Enzymatic formation of beta-cyanoalanine from cyanide. Nature. 1965 Dec 18;208(5016):1207–1208. doi: 10.1038/2081207a0. [DOI] [PubMed] [Google Scholar]
  8. Gros C., Labouesse B. Study of the dansylation reaction of amino acids, peptides and proteins. Eur J Biochem. 1969 Feb;7(4):463–470. doi: 10.1111/j.1432-1033.1969.tb19632.x. [DOI] [PubMed] [Google Scholar]
  9. Hendrickson H. R., Conn E. E. Cyanide metabolism in higher plants. IV. Purification and properties of the beta-cyanolanine synthase of blue lupine. J Biol Chem. 1969 May 25;244(10):2632–2640. [PubMed] [Google Scholar]
  10. 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]
  11. Ressler C., Abe O., Kondo Y., Cottrell B., Abe K. Purification and characterization from Chromobacterium violaceum of an enzyme catalyzing the synthesis of gamma-cyano-alpha-aminobutyric acid and thiocyanate. Biochemistry. 1973 Dec 18;12(26):5369–5377. doi: 10.1021/bi00750a021. [DOI] [PubMed] [Google Scholar]
  12. Strobel G. A. 4-amino-4-cyanobutyric acid as an intermediate in glutamate biosynthesis. J Biol Chem. 1967 Jul 25;242(14):3265–3269. [PubMed] [Google Scholar]
  13. Tolosa E. A., Chepurnova N. K., Khomutov R. M., Severin E. S. Reactions catalysed by cysteine lyase from the yolk sac of chicken embryo. Biochim Biophys Acta. 1969 Feb 11;171(2):369–371. doi: 10.1016/0005-2744(69)90174-0. [DOI] [PubMed] [Google Scholar]
  14. Vesterberg O., Wadström T., Vesterberg K., Svensson H., Malmgren B. Studies on extracellular PROTEINS FROM Staphylococcus aureus. I. Separation and characterization of enzymes and toxins by isoelectric focusing. Biochim Biophys Acta. 1967 Apr 11;133(3):435–445. doi: 10.1016/0005-2795(67)90547-8. [DOI] [PubMed] [Google Scholar]
  15. WADA H., SNELL E. E. The enzymatic oxidation of pyridoxine and pyridoxamine phosphates. J Biol Chem. 1961 Jul;236:2089–2095. [PubMed] [Google Scholar]

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