Selection of novel structural zinc sites from a random peptide library

FEBS Letters

Matsubara, Teruhiko²  Kawahito, Osamu¹  Hiura, Yuko¹  Kawashiro, Katsuhiro¹  Yasuzawa, Mikito¹ 

[1] Department of Chemical Science and Technology, The University of Tokushima, 2-1 Minamijosanjima, Tokushima 770-8506, Japan;Department of Biosciences and Informatics, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522, Japan

关键词: Structural zinc;    Random peptide library;    Metalloenzyme;    Phage display system;    Sequence homology;    ELISA;    enzyme-linked immunosorbent assay;    BSA;    bovine serum albumin;    PBS;    phosphate-buffered saline;    Fmoc;    9-fluorenylmethoxycarbonyl;    TFA;    trifluoroacetic acid;    IC50;    50% inhibitory concentration;    CD;    circular dichroism;    PDB;    Protein Data Bank;    SOD;    superoxide dismutase;   

DOI  :  10.1016/S0014-5793(03)01266-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Zinc ion (Zn²⁺) can be coordinated with four or three amino acid residues to stabilize a protein's structure or to form a catalytic active center. We used phage display selection of a dodecamer random peptide library with Zn²⁺ to identify structural zinc sites. The binding specificity for Zn²⁺ of selected sequences was confirmed using enzyme-linked immunosorbent and competitive inhibition assays. Circular dichroism spectra indicated that the interaction with Zn²⁺ induced a change in conformation, which means the peptide acts as a structural zinc site. Furthermore, a search of protein databases revealed that two selected sequences corresponded to parts of natural zinc sites of copper/zinc superoxide dismutase and zinc-containing ferredoxin. We demonstrated that Zn²⁺-binding sequences selected from the random combinatorial library would be candidates for artificial structural zinc sites.

【 授权许可】

Unknown