Analysis of Molecular Interactions in Heat-induced Aggregation of a Non-inhibitory Serpin Ovalbumin Using a Molecular Chaperone(Biochemistry & Molecular Biology)
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概要
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Aggregation occurs through hydrophobic interactions when a polypeptide chain refolds in non-native states or when genetic variants of biologically active proteins assume inappropriate conformations, as observed in the case of dysfunctional serpins. Here, using the molecular chaperone BiP from bovine liver microsomes, we characterized the hydrophobic nature of the peptide segment which is considered to be a site required for aggregation among a non-inhibitory serpin ovalbumin in a heat-denatured state. Screening of the peptide scan for binding of BiP showed that BiP-binding sites are mostly buried in the folded ovalbumin. When ovalbumin was heat-denatured, the denatured protein was recognized by the antibody that reacts with the hydrophobic surface of the amino-terminal segment of ovalbumin. This antibody significantly suppressed the binding of BiP to denatured ovalbumin. BiP also bound the immobilized peptide in an ATP-dependent manner and the peptide stimulated the ATPase activity of BiP with a K_m of 165 μM and a V_<max> of 0.4nmol/min per milligram. Measurement of surface plasmon resonance showed that the peptide had a K_d of 0.52μM by BiP, lower than that for RCMLA (K_d = 1.1μM) and even lower than that of the peptide P10K, PLSRTLSVAAKK, (K_d = 21μM). These results demonstrate that the aggregation-prone site on heat-denatured ovalbumin has almost the same hydrophobic nature of interacting with the molecular chaperone BiP as the conventionally known peptides that bind to the Escherichia coli chaperone DnaK.
- 社団法人日本農芸化学会の論文
- 2003-05-23
著者
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Kitabatake Naofumi
Research Institute Of Food Science Kyoto University
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Tani Fumito
Department Of Food Science And Technology Faculty Of Agriculture Kyoto University
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Tani Fumito
Research Institute For Food Science Kyoto University
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Tani F
Institute For Fundamental Research Of Organic Chemistry Kyushu University
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Fumito Tani
Division Of Food Science And Biotechnology Graduate School Of Agriculture Kyoto University
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Tani F
Institute For Materials Chemistry And Engineering Kyushu University
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Tani Fumito
Laboratory Of Food And Environmental Sciences Division Of Food Science And Biotechnology Graduate Sc
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Tani Fumito
Institute For Materials Chemistry And Engineering Kyushu University
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Nakanishi Yoko
Division Of Science Of Biological Resources Graduate School Of Science And Technology Kobe Universit
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SHIRAI Nobuaki
Industrial Research Center of Shiga Prefecture
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NAKANISHI Yukiko
Laboratory of Food and Environmental Sciences, Division of Food Science and Biotechnology, Graduate
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KITABATAKE Naofumi
Laboratory of Food and Environmental Sciences, Division of Food Science and Biotechnology, Graduate
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Nakanishi Y
Division Of Food Science And Biotechnology Graduate School Of Agriculture Kyoto University
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Kitabatake N
Division Of Food Science And Biotechnology Graduate School Of Agriculture Kyoto University
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