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Analysis of Leaf Proteins by Two-Dimensional Gel Electrophoresis: Protease Action as Exemplified by Ribulose Bisphosphate Carboxylase/ Oxygenase Degradation and Procedure to Avoid Proteolysis during Extraction.

A previous genetic analysis by two-dimensional electrophoresis of wheat leaf proteins led us to hypothesize that several polypeptides were degradation products of the large subunit of ribulose bisphosphate carboxylase/oxygenase (EC 4.1.1.39).VARIOUS EXTRACTION PROCEDURES INCLUDING THE USE OF PROTEINASE INHIBITORS ALLOWED US TO: (a) confirm that the suspected polypeptides are indeed degradation products; (b) find out under which conditions the proteinases act; (c) find a method which prevents degradation, by boiling the extract in the presence of sodium dodecyl sulfate and 2-mercaptoethanol.