Identification of a Cross-Linked Peptide of a Covalent Complex between Adrenodoxin Reductase and Adrenodoxin.
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- Hara Takayuki
- Section of Biochemistry, Graduate School of Health and Nutrition Sciences, Nakamura Gakuen College
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- Miyata Toshiyuki
- Department of Biology, Faculty of Science, Kyushu University
抄録
A cross-linked complex between bovine NADPH-adrenodoxin reductase (AR) and adrenodoxin (AD) was prepared with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and purified, as described previously [Hara, T. & Kimura, T. (1989) J. Biochem. 105, 594-600]. The covalent complex was S-pyridylethylated and digested with lysylendopeptidase, and the resulting peptides were separated by reversed-phase HPLC to identify the cross-linked peptide. Comparison of the HPLC chromatograms of the peptides showed that (i) two tandem peptides (K-4 and K-5) from AD and a peptide (K-1) from AR were missing in the chromatogram of the peptides of the covalent complex and (ii) a single new peak was observed in the chromatogram of the peptides from the covalent complex. Amino acid composition and sequence analyses showed that the newly observed peptide was a covalently cross-linked peptide formed between a peptide K-4-K-5 (Ile-25-Lys-98) derived from AD and a peptide K-1 (Ser-l-Lys-27) derived from AR, in which an amide bond had been formed between the ε -amino group of Lys-66 in AD and the γ-carboxyl group of Glu-4 in AR. These results indicate that the binding site of AR with AD is localized in the amino-terminal part of AR and that of AD with AR is localized around Lys-66 of AD. The six clustered basic amino acid residues (His-24, Lys-27, His-28, His-29, Arg-31, and His-33) present in the amino-terminal portion of AR and the eight clustered acidic amino acid residues (Glu-65, Glu-68, Asp-72, Glu-73, Glu-74, Asp-76, Asp-79, and Asp-86) present in the middle part of AD may play an important role in the complex formation.
収録刊行物
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- The Journal of Biochemistry
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The Journal of Biochemistry 110 (2), 261-266, 1991
The Japanese Biochemical Society
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詳細情報 詳細情報について
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- CRID
- 1570572703173612160
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- NII論文ID
- 130003531469
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- ISSN
- 0021924X
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- 本文言語コード
- en
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- データソース種別
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- CiNii Articles