TY - JOUR
T1 - Selection of ganglioside GM1-binding peptides by using a phage library
AU - Matsubara, Teruhiko
AU - Ishikawa, Dai
AU - Taki, Takao
AU - Okahata, Yoshio
AU - Sato, Toshinori
N1 - Funding Information:
This work is partially supported by Grants-in-Aid for Scientific Research number 09240211 and 10134213 (T.S.) from the Ministry of Education, Science, Sports and Culture of Japan.
PY - 1999/8/6
Y1 - 1999/8/6
N2 - 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 IC50 of 24, 13 and 1.0 μM, respectively. These peptides will be useful for searching functional roles of ganglioside GM1. Copyright (C) 1999 Federation of European Biochemical Societies.
AB - 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 IC50 of 24, 13 and 1.0 μM, respectively. These peptides will be useful for searching functional roles of ganglioside GM1. Copyright (C) 1999 Federation of European Biochemical Societies.
KW - Carbohydrate recognition
KW - Galβ1→3GalNAcβ1→4(NeuAcα2→3)Galβ1→4Glcβ1→1'Cer
KW - Ganglioside
KW - Monolayer
KW - Phage-displayed peptide library
KW - Quartz-crystal microbalance
UR - http://www.scopus.com/inward/record.url?scp=0032806603&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032806603&partnerID=8YFLogxK
U2 - 10.1016/S0014-5793(99)00962-X
DO - 10.1016/S0014-5793(99)00962-X
M3 - Article
C2 - 10456319
AN - SCOPUS:0032806603
SN - 0014-5793
VL - 456
SP - 253
EP - 256
JO - FEBS Letters
JF - FEBS Letters
IS - 2
ER -
