JXB Advance Access originally published online on August 27, 2004
Journal of Experimental Botany 2004 55(406):2241-2249; doi:10.1093/jxb/erh247
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RESEARCH PAPER |
A comparative study of the role of the major proteinases of germinated common bean (Phaseolus vulgaris L.) and soybean (Glycine max (L.) Merrill) seeds in the degradation of their storage proteins
1Laboratory of Protein Chemistry, Moldova State University, Mateevici str.60, MD-2009, Kishinev, Republic of Moldov
2Department of Biological Sciences, State University of New York at Binghamton, Binghamton, NY 13902-6000, USA
* To whom correspondence should be addressed. Fax: +37 322 244 248. E-mail: vaintrau{at}usm.md
Two types of cysteine proteases, low-specificity enzymes from the papain family and Asn-specific from the legumain family are generally considered to be the major endopeptidases responsible for the degradation of seed storage proteins during early seedling growth. The action of the corresponding enzymes (CPPh1 and LLP, respectively) from common bean (Phaseolus vulgaris L.) on phaseolin (the common bean storage protein), and on the homologous soybean (Glycine max (L.) Merrill) storage protein, ß-conglycinin, was studied. Under the action of LLP, proteolysis of phaseolin was limited to cleavage of its interdomain linker. No cleavage of the interdomain linker occurred in ß-conglycinin with LLP. LLP action was restricted to splitting off the disordered N-terminal extensions of 
and ' subunits. No extensive hydrolysis (degradation to short TCA-soluble peptides) of either protein occurred under the action of LLP. CPPh1 cleaved the phaseolin subunits into roughly half-sized fragments at the onset of proteolysis. The cleavage was accompanied by a small (8–10%) decrease of protein. No decrease of protein occurred with further incubation. Thus the two most active proteinases detected in common bean seedlings individually were incapable of the extensive degradation of phaseolin. Extensive hydrolysis of phaseolin was only achieved by the consecutive action of LLP and CPPh1. Similar cleavages occurred during the action of CPPh1 on ß-conglycinin. However, by contrast with phaseolin, CPPh1 by itself accomplished the extensive hydrolysis of ß-conglycinin. The differences in the course of proteolysis of the proteins studied were determined by their structural peculiarities.
Key words: ß-conglycinin, legumain-like proteinase, papain-like proteinase, phaseolin, proteolysis, seed storage protein degradation