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Purification of Glyoxysomal Catalase and Immunochemical Comparison of Glyoxysomal and Leaf Peroxisomal Catalase in Germinating Pumpkin Cotyledons

Junji Yamaguchi, Mikio Nishimura
Junji Yamaguchi
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Mikio Nishimura
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Published February 1984. DOI: https://doi.org/10.1104/pp.74.2.261

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Abstract

As a step to study the mechanism of the microbody transition (glyoxysomes to leaf peroxisomes) in pumpkin (Cucurbita sp. Amakuri Nankin) cotyledons, catalase was purified from glyoxysomes. The molecular weight of the purified catalase was determined to be 230,000 to 250,000 daltons. The enzyme was judged to consist of four identical pieces of the monomeric subunit with molecular weight of 55,000 daltons. Absorption spectrum of the catalase molecule gave two major peaks at 280 and 405 nanometers, showing that the pumpkin enzyme contains heme. The ratio of absorption at 405 and 280 nanometers was 1.0, the value being lower than that obtained for catalase from other plant sources. These results indicate that the pumpkin glyoxysomal catalase contains the higher content of heme in comparison with other plant catalase.

The immunochemical resemblance between glyoxysomal and leaf peroxisomal catalase was examined by using the antiserum specific against the purified enzyme preparation from pumpkin glyoxysomes. Ouchterlony double diffusion and immunoelectrophoretic analysis demonstrated that catalase from both types of microbodies cross-reacted completely whereas the immunotitration analysis showed that the specific activity of the glyoxysomal catalase was 2.5-fold higher than that of leaf peroxisomal catalase. Single radial immunodiffusion analysis showed that the specific activity of catalase decreased during the greening of pumpkin cotyledons.

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Purification of Glyoxysomal Catalase and Immunochemical Comparison of Glyoxysomal and Leaf Peroxisomal Catalase in Germinating Pumpkin Cotyledons
Junji Yamaguchi, Mikio Nishimura
Plant Physiology Feb 1984, 74 (2) 261-267; DOI: 10.1104/pp.74.2.261

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Purification of Glyoxysomal Catalase and Immunochemical Comparison of Glyoxysomal and Leaf Peroxisomal Catalase in Germinating Pumpkin Cotyledons
Junji Yamaguchi, Mikio Nishimura
Plant Physiology Feb 1984, 74 (2) 261-267; DOI: 10.1104/pp.74.2.261
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Plant Physiology
Vol. 74, Issue 2
February 1984
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