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. 1976 May;57(5):795–798. doi: 10.1104/pp.57.5.795

Partial Purification and Characterization of Two Peptide Hydrolases from Pea Seeds

J Bruce Caldwell 1, Lindsay G Sparrow 1
PMCID: PMC542119  PMID: 16659571

Abstract

Two peptide hydrolases have been found in pea seeds (Pisum sativum var. Greenfeast) and extensively purified by ion exchange chromatography using benzoyl-dl-arginine-p-nitroanilide as substrate. The enzymes which both have molecular weights of 65,000 can be separated by anion exchange chromatography but are otherwise virtually identical in the properties tested. They did not hydrolyze several common protease substrates but readily hydrolyzed small peptides containing basic amino acids on the carboxyl side of these residues. They are completely inhibited by diisopropylfluorophosphate and are inhibited to varying extents by thiol reagents.

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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. Estimation of the molecular weights of proteins by Sephadex gel-filtration. Biochem J. 1964 May;91(2):222–233. doi: 10.1042/bj0910222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. CRESTFIELD A. M., MOORE S., STEIN W. H. The preparation and enzymatic hydrolysis of reduced and S-carboxymethylated proteins. J Biol Chem. 1963 Feb;238:622–627. [PubMed] [Google Scholar]
  3. Cameron E. C., Mazelis M. A Nonproteolytic "Trypsin-like" Enzyme: Purification and Properties of Arachain. Plant Physiol. 1971 Sep;48(3):278–281. doi: 10.1104/pp.48.3.278. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Catsimpoolas N., Funk S. K., Wang J., Kenney J. Isoelectric fractionation and some properties of a protease from soyabean seeds. J Sci Food Agric. 1971 Feb;22(2):79–82. doi: 10.1002/jsfa.2740220209. [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. ERLANGER B. F., KOKOWSKY N., COHEN W. The preparation and properties of two new chromogenic substrates of trypsin. Arch Biochem Biophys. 1961 Nov;95:271–278. doi: 10.1016/0003-9861(61)90145-x. [DOI] [PubMed] [Google Scholar]
  7. Elleman T. C. Aminopeptides of pea. Biochem J. 1974 Jul;141(1):113–118. doi: 10.1042/bj1410113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gleeson P., Sparrow L. G., Ward C. W. Aryl acylamidase activity in larvae of the webbing clothes moth, Tineola bisselliella. Comp Biochem Physiol B. 1976;53(2):187–190. doi: 10.1016/0305-0491(76)90033-x. [DOI] [PubMed] [Google Scholar]
  9. Hayashi R., Hata T. Action of yeast proteinase C on synthetic peptides and poly- ,L-amino acids. Biochim Biophys Acta. 1972 May 18;263(3):673–679. doi: 10.1016/0005-2795(72)90049-9. [DOI] [PubMed] [Google Scholar]
  10. Hoagland R. E., Graf G. The purification and properties of an amidohydrolase from soybean. Can J Biochem. 1974 Oct;52(10):903–910. doi: 10.1139/o74-127. [DOI] [PubMed] [Google Scholar]
  11. Jones S. R., Hofmann T. Penicillocarboxypeptidase-S, a nonspecific SH-dependent exopeptidase. Can J Biochem. 1972 Dec;50(12):1297–1310. doi: 10.1139/o72-175. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Marks N., Datta R. K., Lajtha A. Partial resolution of brain arylamidases and aminopeptidases. J Biol Chem. 1968 Jun 10;243(11):2882–2889. [PubMed] [Google Scholar]
  14. NAUGHTON M. A., SANGER F., HARTLEY B. S., SHAW D. C. The amino acid sequence around the reactive serine residue of some proteolytic enzymes. Biochem J. 1960 Oct;77:149–163. doi: 10.1042/bj0770149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. ROSEN H. A modified ninhydrin colorimetric analysis for amino acids. Arch Biochem Biophys. 1957 Mar;67(1):10–15. doi: 10.1016/0003-9861(57)90241-2. [DOI] [PubMed] [Google Scholar]
  16. Visuri K., Mikola J., Enari T. M. Isolation and partial characterization of a carboxypeptidase from barley. Eur J Biochem. 1969 Jan;7(2):193–199. doi: 10.1111/j.1432-1033.1969.tb19591.x. [DOI] [PubMed] [Google Scholar]
  17. Ward C. W. Spectral data for modified assay of benzoyl-arginine p-nitroanilide-hydrolysing enzymes in crude extracts. Anal Biochem. 1973 Dec;56(2):596–600. doi: 10.1016/0003-2697(73)90228-5. [DOI] [PubMed] [Google Scholar]
  18. Zemchik E. I., Shutov A. D., Vaintraub I. A. Chastichnaia ochistka i kharakteristika arilamidazy iz semian viki. Biokhimiia. 1973 Sep-Oct;38(5):964–970. [PubMed] [Google Scholar]

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