Selection of ganglioside GM1‐binding peptides by using a phage library

FEBS Letters

Matsubara, Teruhiko²  Sato, Toshinori²  Taki, Takao¹  Okahata, Yoshio²  Ishikawa, Dai¹ 

[1]Cellular Technology Institute, Otsuka Pharmaceutical, 463-10 Kagasuno, Kawauchi, Tokushima 771-0192, Japan

[2]Department of Biomolecular Engineering, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan

关键词: Ganglioside;    Galβ1→3GalNAcβ1→4(NeuAcα2→3)Galβ1→4Glcβ1→1'Cer;    Carbohydrate recognition;    Monolayer;    Phage-displayed peptide library;    Quartz-crystal microbalance;    Cer;    ceramide;    Gal;    galactose;    NeuAc;    N-acetylneuraminic acid;    GM1;    Galβ1→3GalNAcβ1→4(NeuAcα2→3)Galβ1→4Glcβ1→1'Cer;    ELISA;    enzyme-linked immunosorbent assay;    BSA;    bovine serum albumin;    TBS;    Tris-buffered saline;    QCM;    quartz-crystal microbalance;    TU;    transducing units;    CTB;    cholera toxin B subunit;   

DOI  :  10.1016/S0014-5793(99)00962-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Ganglioside Galβ1→3GalNAcβ1→4(NeuAcα2→3)Galβ1→4Glcβ1→1'Cer (GM1)-binding peptides were obtained from a phage-displayed pentadecapeptide library by an affinity selection. The selection processes were in situ-monitored by a quartz-crystal microbalance method, on which a ganglioside GM1 monolayer was transferred. After five rounds of biopanning, the DNA sequencing of 18 selected phages showed that only three individual clones were selected. The peptide sequences of the random region were found to be DFRRLPGAFWQLRQP, GWWYKGRARPVSAVA and VWRLLAPPFSNRLLP. Binding constants of these phage clones to the GM1 monolayer were 10¹⁰ M⁻¹. Three synthetic pentadecapeptides inhibited the binding of cholera toxin B subunit to the GM1 monolayer with an IC ₅₀ of 24, 13 and 1.0 μM, respectively. These peptides will be useful for searching functional roles of ganglioside GM1.

【 授权许可】

Unknown