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Soybean subtilisin-like protease involved in initiating storage protein degradation

Published online by Cambridge University Press:  22 February 2007

Xiaowen Liu ,
Zheng Zhang ,
Neel Barnaby ,
Karl A. Wilsona  and
Anna Tan-Wilsona
Xiaowen Liu
Affiliation:
Department of Biological Sciences, State University of New York at Binghamton, Binghamton, NY, 13902-6000, USA
Zheng Zhang
Affiliation:
Department of Biological Sciences, State University of New York at Binghamton, Binghamton, NY, 13902-6000, USA
Neel Barnaby
Affiliation:
Department of Biological Sciences, State University of New York at Binghamton, Binghamton, NY, 13902-6000, USA
Karl A. Wilsona
Affiliation:
Department of Biological Sciences, State University of New York at Binghamton, Binghamton, NY, 13902-6000, USA
Anna Tan-Wilsona*
Affiliation:
Department of Biological Sciences, State University of New York at Binghamton, Binghamton, NY, 13902-6000, USA
*
*Correspondence Fax: 607-777-6521 Email: annatan@binghamton.edu
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Abstract

Abstract A partial cDNA fragment (1.1kb) of protease C1, an enzyme that initiates the proteolytic degradation of the β-conglycinin storage proteins in the soybean (Glycine max [L.] Merrill. cv. Amsoy 71) through limited action at Glurich regions, has been cloned by reverse transcription-polymerase chain reaction (RT-PCR). The sequence was extended toward the 5' end by another 314 bases. The nucleotide sequence shows that protease C1 belongs to the subtilisin family of serine proteases. The sequence encompasses the critical Asp, His and Ser residues of the catalytic triad, as well as the Asn at the binding site. Northern analysis shows the presence of a 2.5kb mRNA not only in seedling cotyledons, but also in developing seeds. The developing seeds, and even dry seeds, exhibit phenylmethylsulfonyl fluoride (PMSF) and EDTA-sensitive protease activity that only cleaves the α- and α'-, but not the β-subunits, of soybean β-conglycinin. The discrete proteolytic intermediates produced are of the same sizes as those produced by pure enzyme, as are the final 50 kDa and 48 kDa products. The activity is also sensitive to inhibition by synthetic poly-L-Glu, all characteristics of purified protease C1. These data suggest that protease C1, or an enzyme very similar to it, is synthesized in a form that is active in vitro. Because seeds do accumulate β-conglycinin and because there is very little evidence of the proteolytic products of protease C1 action in extracts of dry seeds, one can surmise that the protease C1 is not particularly active in vivo during seed maturation. GenBank lists the sequence of the 1.1kb fragment with accession number AAD02075.

Keywords

protease C1subtilisin-like proteaseGlycine maxseed developmentseedling cotyledonssoybeanstorage protein mobilization
Type
Research Article
Information
Seed Science Research , Volume 11 , Issue 1 , March 2001 , pp. 55 - 68
Copyright
Copyright © Cambridge University Press 2001

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