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Characterization of carboxypeptidase I of mung bean seeds

Published online by Cambridge University Press:  19 September 2008

Karl A. Wilson ,
Mary Russell ,
John F. Quackenbush  and
Anna L. Tan-Wilson
Karl A. Wilson*
Affiliation:
Department of Biological Sciences, State University of New York at Binghamton, Binghamton, NY 13902–6000, USA
Mary Russell
Affiliation:
Department of Biological Sciences, State University of New York at Binghamton, Binghamton, NY 13902–6000, USA
John F. Quackenbush
Affiliation:
Department of Biological Sciences, State University of New York at Binghamton, Binghamton, NY 13902–6000, USA
Anna L. Tan-Wilson
Affiliation:
Department of Biological Sciences, State University of New York at Binghamton, Binghamton, NY 13902–6000, USA
*
*Correrpondence
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Abstract

There is a carboxypeptidase in mung bean seeds that is localized in the protein bodies, the same vacuoles in which seed storage proteins are sequestered. This carboxypeptidase, called carboxypeptidase I (or Cpase I) has been purified by a series of ion-exchange and gel filtration columns. The pure enzyme consists of a single polypeptide chain with a MW of 41700 by SDS-PAGE or 42000 by size-exclusion HPLC. It has a pl of 4.36 and is a serine carboxypeptidase as shown by its inactivation by phenylmethylsulfonyl fluoride, and its resistance to other proteolytic inhibitory reagents. A survey of its activity with Cbz-dipeptides shows preference for C-terminal amino acids that are large, hydrophobic residues, and a small aliphatic amino acid such as alanine, but not glycine, at the penultimate amino acid residue. Cpase I can convert a trypsin inhibitor of the mung bean to its proteolytic intermediate lacking four amino acid residues at its C-terminus. This proteolytic intermediate is detected in the mung bean cotyledons during early growth. Levels of both the activity and immunological cross-reacting forms of this enzyme start high and decrease during early growth.

Keywords

carboxypeptidasegerminationmung beanproteolysisdegradationtrypsin inhibitor
Type
Physiology and Biochemistry
Information
Seed Science Research , Volume 5 , Issue 4 , December 1995 , pp. 209 - 218
Copyright
Copyright © Cambridge University Press 1995

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References

Baulcombe, D.C., Barker, R.F. and Jarvis, M.G. (1987) Agibberellin-responsive wheat gene has homology to yeast carboxypeptidase Y. Journal of Biological Chemistry 262, 13726–13735.CrossRefGoogle ScholarPubMed
Baumgartner, B. and Chrispeels, M.J. (1977) Purification and characterization of vicilin peptidohydrolase, the major endopeptidase in the cotyledons of mung bean seedings. European Journal of Biochemistry 77, 223–233.CrossRefGoogle Scholar
Bayer, E.A., Ben-Hur, H., and Wilchek, M. (1990) Analysis of proteins and glycoproteins on blots. Methods in Enzymology 184, 415–429.CrossRefGoogle ScholarPubMed
Bradley, D. (1992) Isolation and characterization of a gene encoding a carboxypeptidase Y-like protein from Ara-bidopsis thaliana. Plant Physiology 98, 1526–1529.CrossRefGoogle ScholarPubMed
Breddam, K. (1986) Serine Carboxypeptidases. A review. Carlsberg Research Communications 51, 83–128.CrossRefGoogle Scholar
Breddam, K. and Sorensen, S.B. (1987) Isolation of carboxypeptidase III from malted barley by affinity chromatography. Carlsberg Research Communications 52, 275–283.CrossRefGoogle Scholar
Breddam, K., Sorensen, S.B. and Svendsen, I. (1987) Primary structure and enzymatic properties of carboxypeptidase II from wheat germ. Carlsberg Research Communications 52, 297–311.CrossRefGoogle Scholar
Chrispeels, M.J., Baumgartner, B. and Harris, N. (1976) Regulation of reserve protein metabolism in the cotyledons of mung bean seedlings. Proceedings of the National Academy of Sciences, USA 73, 3168–3172.CrossRefGoogle ScholarPubMed
Davis, B.J. (1964) Disc electrophoresis II: Method and application to human serum proteins. Annals of the New York Academy of Sciences 121, 404–427.CrossRefGoogle ScholarPubMed
Duarte, I.C., Ricardo, C.P.P. and Duque-Magalhaes, M.C. (1993) Peptide hydrolases in cotyledons of germinating lupin. Phytochemistry 33, 35–40.CrossRefGoogle Scholar
Doi, E., Komori, N., Matoba, T. and Morita, Y. (1980a) Some properties of carboxypeptidases in germinating rice seeds and rice leaves. Agricultural and Biological Chemistry 44, 77–81.Google Scholar
Doi, E., Komori, N., Matoba, T. and Morita, Y. (1980b) Purification and some properties of a carboxypeptidase in rice bran. Agricultural and Biological Chemistry 44, 85–92.Google Scholar
Ihle, J.N. and Dure, L.S. (1972) The developmental biochemistry of cottonseed embryogenesis and germination. Journal of Biological Chemistry 247, 5041–5047.CrossRefGoogle ScholarPubMed
Kubota, Y., Shoji, S., Yamanaka, T. and Yamato, M. (1976) Carboxypeptidases from germinating soybeans. I. Purification and properties of two carboxypeptidases. Yakugaku Zasshi 96, 639–647.CrossRefGoogle ScholarPubMed
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.CrossRefGoogle ScholarPubMed
Preston, K.R. and Kruger, J.E. (1976) Purification and properties of two proteolytic enzymes with carboxypeptidase activity in germinated wheat. Plant Physiology 58, 516–520.CrossRefGoogle ScholarPubMed
Scalet, M. and Alpi, A. (1985) Partial purification and characterization of several carboxypeptidase-like activities present in leaf extracts of Medicago sativa. Journal of Plant Physiology 121, 47–57.CrossRefGoogle Scholar
Shutov, A.D. and Vaintraub, I.A. (1987) Degradation of storage proteins in germinating seeds. Phytochemistry 26, 1557–1566.CrossRefGoogle Scholar
Sorensen, S.B., Breddam, K. and Svendsen, I.B. (1986) Primary structure of carboxypeptidase I from malted barley. Carlsberg Research Communications 51, 475–485.CrossRefGoogle Scholar
Sorensen, S.B., Svendsen, I. and Breddam, K. (1987) Primary structure of carboxypeptidase II from malted barley. Carlsberg Research Communications 52, 285–295.CrossRefGoogle Scholar
Sorensen, S.B., Svendsen, I. and Breddam, K. (1989) Primary structure of carboxypeptidase III from malted barley. Carlsberg Research Communications 54, 193–202.CrossRefGoogle ScholarPubMed
Towbin, H., Staelin, T. and Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, USA 76, 4350–4354.CrossRefGoogle ScholarPubMed
Wilson, K.A. (1986) Role of proteolytic enzymes in the mobilization of protein reserves in the germinating dicot seeds. pp 19–47 in Dalling, M.J. (Ed.) Plant proteolytic enzymes. Boca Raton, Florida, CRC Press.Google Scholar
Wilson, K.A. and Chen, J.C. (1983) Amino acid sequence of mung bean trypsin inhibitor and its modified forms appearing during germination. Plant Physiology 71, 341–349.CrossRefGoogle ScholarPubMed
Wilson, K.A. and Tan-Wilson, A.L. (1987) Characterization of the proteinase that initiates the degradation of the trypsin inhibitor in germinating mung beans (Vigna radiata). Plant Physiology 84, 93–98.CrossRefGoogle ScholarPubMed
Wilson, K.A., Rightmire, B.R. and Tan-Wilson, A.L. (1985) Involvement of carboxypeptidase in the degradation of the mung bean (Vigna radiata) trypsin inhibitor during germination and early seedling growth. Qualitas Plantarum Foods Human Nutrition 35, 195–211.CrossRefGoogle Scholar
Wray, W., Boulikas, T., Wray, V.P. and Hancock, R. (1981) Silver staining of proteins in polyacrylamide gels. Analytical Biochemistry 118, 197–208.CrossRefGoogle ScholarPubMed
Yamaoka, Y., Ohba, Y., Takeuchi, M. and Morohashi, Y. (1993) Isolation and properties of a carboxypeptidase from cotyledons of germinated mung bean seeds. Plant Science 95, 1–7.CrossRefGoogle Scholar