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Selection of ganglioside GM1‐binding peptides by using a phage library
Author(s) -
Matsubara Teruhiko,
Ishikawa Dai,
Taki Takao,
Okahata Yoshio,
Sato Toshinori
Publication year - 1999
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(99)00962-x
Subject(s) - biopanning , ganglioside , phage display , peptide library , peptide , chemistry , microbiology and biotechnology , binding site , binding selectivity , cholera toxin , dna , monolayer , biochemistry , stereochemistry , biology , peptide sequence , gene
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 10 10 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.