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

利用二维凝胶电泳分析叶片蛋白质:以核酮糖二磷酸羧化酶/加氧酶降解为例阐述蛋白酶的作用以及避免提取过程中蛋白水解的方法

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Abstract

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.

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