Protease inhibitor studies and cloning of a serine carboxypeptidase cDNA from germinating seeds of pea (Pisum sativum L.)

Abstract

The nature of the proteolytic activity found within the germinating pea (Pisum sativum) seed, 4 days from the initiation of inhibition, was determined by the use of specific protease inhibitors. These studies have shown most of the activity to belong to metallo or metal-activated and serine proteases. In order to investigate further the serine protease activity, a pea cotyledon germination cDNA library was, therefore, screened with a wheat cDNA (2437) [Baulcombe, D. C., Barker, R. E and Jarvis, M. G. (1987) J. Biol. Chem. 262, 13726-13735] which had extensive similarity to the yeast serine carboxypeptidase Y gene. A positive cDNA clone (pNY551) was obtained which had extensive similarity to the four carboxypeptidases, Arabidopsis thaliana carboxypeptidase Y-like protein, rice serine carboxypeptidase III, barley serine carboxypeptidase III and wheat serine carboxylpeptidase III precursor. Northern-blot analysis showed mRNA homologous to pNY551 to be expressed in late developmental pea seed and again during germination.