Open AccessPurification and characterization of heme‐containing low‐activity form of catalase from greening pumpkin cotyledons
Author(s) -
YAMAGUCHI Junji,
NISHIMURA Mikio,
AKAZAWA Takashi
Publication year - 1986
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1986.tb09870.x
Subject(s) - catalase , molecular mass , heme , biochemistry , greening , peroxisome , chemistry , enzyme , microbody , peroxidase , molecule , biology , organic chemistry , receptor , ecology
In germinating pumpkin seeds, catalase is synthesized as a precursor (59‐kDa) form, with molecular mass larger than the mature molecule (55 kDa). Both the precursor and mature forms of catalase are localized in the microbodies, i.e., glyoxysomes and leaf peroxisomes [ Proc. Natl. Acad. Sci. USA 81 , 4809–4813 (1984)]. We have now purified the 59‐kDa catalase precursor and compared its properties with those of the 55‐kDa mature molecule. The molar catalytic activity of the 59‐kDa catalase was tenfold lower than that of the 55‐kDa molecule, whereas the heme content was found to be same, with both forms containing four hematin groups per molecule. It is inferred from these results that the low activity of the 59‐kDa molecule is not related to the binding of heme to the protein, but presumably involves conformational differences between the 59‐kDa and 55‐kDa molecules. We have further found that the reduction of total catalase activity in pumpkin cotyledons during greening was due to a decrease in the amount of the enzymically active 55‐kDa catalase accompanying an increase in the 59‐kDa molecule.