Protein rotation study of cytochrome P-450 in submitochondrial particles: Effect of KCl and intermolecular interactions with redox partners.
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概要
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The rotational diffusion of cytochrome P-450 in submitochondrial particles (SMP) of bovine adrenocortical mitochondria was measured by detecting the decay of absorption anisotropy, r(t), after photolysis of the heme•CO complex by a vertically polarized laser flash. Analysis of r(t) was based on a "rotation-about-membrane normal" model. The measurements were used to investigate the effect of KCI on intermolecular interactions involving cytochrome P-450 and to investigate the interactions of cytochrome P-450 with other redox partners. The rotational diffusion of cytochrome P-450 was significantly dependent on KC1 concentration. When the KC1 concentration was increased from 0 to 1, 000 mM, the mobile population of cytochrome P-450 was increased from 33 to 82%. After removing the KCI, the mobile population of cytochrome P-450 returned to the original 33%. These results suggest that nonspecific protein aggregates are dissociated by the presence of KC1, possibly due to the change in electrostatic interactions, resulting in mobilization of cytochrome P-450. SMP were observed to be nearly free from adrenodoxin and adrenodoxin reductase. The addition of adrenodoxin to SMP increased the mobile population of cyto-chrome P-450 from 35 to 54%. Further addition of adrenodoxin reductase to SMP containing adrenodoxin immobilized cytochrome P-450 by 6%. The addition of only adrenodoxin reductase to SMP, however, did not immobilize cytochrome P-450. The present results are consistent with our previous observations [Ohta, Y., Mitani, F., Ishimura, Y., Yanagibashi, K., Kawamura, M., & Kawato, S. (1990) J. Biochem. 107, 97-104] that cholesterol-bearing P-4505scc forms a transient ternary association with adrenodoxin and adrenodoxin reductase.
- 社団法人 日本生化学会の論文
著者
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Kawato Suguru
Institute Of Physics College Of Arts And Sciences The University Of Tokyo At Komaba
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Hara Takayuki
Department Of Biochemistry Graduate School Of Health And Nutrition Sciences Nakamura Gakuen Universi
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Ohta Yoshihiro
Institute Of Physics College Of Arts And Sciences The University Of Tokyo At Komaba
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Kawamura Masahiro
Department Of Pharmacology (1) The Jikei University School Of Medicine
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KAWAMURA Masahiro
Department of Pharmacology, The Jikei University School of Medicine
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KAWATO Suguru
Institute of Physics, College of Arts and Sciences, The University of Tokyo at Komaba
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YANAGIBASHI Kazutoshi
Department of Pharmacology I, The Jikei University School of Medicine
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