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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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