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Proteolysis of ribulose‐1,5‐bisphosphate carboxylase/oxygenase in Citrus leaf extracts
Author(s) -
GarcíaMartínez José L.,
Moreno Joaquín
Publication year - 1986
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1986.tb05938.x
Subject(s) - ribulose 1,5 bisphosphate , pyruvate carboxylase , oxygenase , biochemistry , pepstatin , protein subunit , proteolysis , chemistry , rubisco , gel electrophoresis , citrus × sinensis , polyacrylamide gel electrophoresis , ribulose , biology , enzyme , orange (colour) , protease , food science , gene
Ribulose‐1,5‐bisphosphate carboxylase/oxygenase (RuBP carboxylase, EC 4.1.1.39) has been purified from orange [ Citrus sinensis (L.) Osbeck cv. Washington Navel] leaves using sucrose gradient centrifugation in a fixed angle rotor. Following sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE), two major bands corresponding to the two subunits of RuBP carboxylase were found. The large subunit coincided with the polypeptide band that has been previously reported to be preferentially mobilized during the spring and summer flush periods. The degradation of RuBP carboxylase during autodigestion of Citrus leaf extracts, investigated by SDS‐PAGE, occurred mainly at acidic (2.5‐5.5) pH. The two subunits showed differences in the rate of degradation, the smaller being more rapidly hydrolyzed than the larger. At least four proteolytic activities were identified by means of inhibitor experiments: 1) a pepstatin A‐sensitive activity that acts on both RuBP carboxylase subunits, 2) a mercurial ( p ‐hydroxymercuribenzoate and p ‐chloromercuriphenylsulfonate)‐sensitive activity that degrades only the small subunit, 3) an EDTA‐sensitive activity that hydrolyzes both the large and small subunits, and 4) a mercurial‐stimulated activity that acts only on the large subunit. It is suggested that the last two proteases may be responsible for the degradation of RuBP carboxylase observed in vivo during the periods of mobilization of leaf protein in Citrus .