Preparation of peptides which mimic glycosphingolipids by using phage peptide library and their modulation on β‐galactosidase activity

FEBS Letters

Taki, Takao¹  Handa, Shizuo¹  Hamasaki, Hiroko¹  Ishikawa, Dai¹ 

[1] Department of Biochemistry, School of Medicine, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113, Japan

关键词: Phage-displayed random peptide library;    Glyco-replica peptide;    Glycosidase inhibitor;    Gal;    galactose;    GlcNAc;    N-acetylglucosamine;    Lc3Cer;    GlcNAcβ1→3Galβ1→4Glcβ1→1Cer;    Lc4Cer;    Galβ1→3GlcNAcβ1→3Galβ1→4Glcβ1→1Cer;    nLc4Cer;    Galβ1→4GlcNAcβ1→3Galβ1→4Glcβ1→1Cer;    PBS;    phosphate buffered saline;    TLC;    thin-layer chromatography;   

DOI  :  10.1016/S0014-5793(97)01386-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We describe the use of a phage-displayed random pentadecamer peptide library for searching glycosphingolipid mimicking peptides. Two phage clones (AD-1 and AD-2) were selected by biopanning using monoclonal antibody AD117m, directed to lactotetraosylceramide (Lc₄Cer). The amino acid sequences of the selected clones showed high homology (VPPXFXXXY) in 9-mer. Three phage clones were selected by using monoclonal antibody H11, directed to neolactotetraosylceramide (nLc₄Cer), the linkage isomer of Lc₄Cer, and the displayed amino acid sequences were compared. One of these peptides showed the same amino acid sequence as that of AD-2 except for one amino acid substitution. Pentadecamer, 9-mer and point mutated 9-mer peptides were synthesized on the basis of the displayed amino acid sequences. Binding activity of the peptides to the monoclonal antibodies or Ricinus communis lectin showed that 9-mer peptides are enough to mimic the epitope carbohydrate structure. Furthermore, six of the synthesized peptides inhibited Jack bean β-galactosidase activity towards nLc₄Cer at a high concentration of the enzyme, whereas at lower enzyme concentrations some peptides showed potent activation of the enzyme activity. This is the first report of carbohydrate mimicking peptides which modulate glycosidase activity.

【 授权许可】

Unknown