Interaction of α_2-Macroglobulin with Serine Proteinase from Aspergillus sojae (Biological Chemistry)
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概要
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The interaction of α_2-macroglobulin (α_2M) with a major fraction of serine proteinase (SepII) from Aspergillus sojae was investigated. The stoichiometry of the reaction of α_2M with SepII showed that SepII bound to α_2M in a molar ratio of about 2:1. The complex (α_2M-SepII) was separated by PAGE and chromatograph. About 80% inhibition of the proteinase activity as to the hydrolysis of casein and succinyl-L-leucyl-L-leucyl-L-valyl-L-tyrosyl-7-amino-4-methylcoumarin (Suc-LLVY-MCA) was observed. The residual activity of the complex was completely abolished by chymostatin and antipain, which were powerful inhibitors with Ki values of 3 × 10^<-8> and 5 × 10^<-8>, respectively. Immunoelectrophoresis of the complex showed its weak cross-reactivity with antiserum to SepII (anti-SepII). The activity of the complex was partially abolished by the anti-SepII. Examination of electron micrographs left little doubt that a comformational change in shape took place. The entrapment reaction with α_2M was completed within 30 sec. When the α_2M-SepII complex was incubated at pH 3.6, a triphasic course was observed. Activation of the complex was observed within 10〜17 min, while inactivation was observed after 10〜17 min incubation. And then the activity remained at several % of the initial level. The complex was found to exhibit enhanced pH stability at pH 3.6, while alkali and heat denaturation were observed not to have drastic effects on its stability.
- 社団法人日本農芸化学会の論文
- 1988-09-23
著者
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ICHISHIMA EIJI
Department of Bioengineering, Graduate School of Engineering, Soka University
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NAKAJIMA TASUKU
Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University
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Ichishima Eiji
Department Of Agricultural Chemistry Faculty Of Agriculture Tohoku University
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Iida H
Laboratory Of Enzymology And Microbial Chemistry Tokyo University Of Agriculture And Technology
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Iida Hiroshi
Laboratory Of Zoology Graduate School Of Agriculture Kyushu University
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Nakajima Tasuku
Department Of Agricultural Chemistry Faculty Of Agriculture Tohoku University
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Iida Hiroshi
Laboratory Of Enzymology And Microbial Chemistry Department Of Agricultural Chemistry Tokyo Universi
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ICHISHIMA Eiji
Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University
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