Selection of sugar-binding peptides from a scaffold-containing library

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Molecular evolution of binding affinity of artificial peptides by random mutation and affinity selection was performed. DNA fragments encoding ganglioside Galβ1-3GalNAcβ1-4(Neu5Acα2-3)Galβl-4Glcβ1- 1′Cer (GM1)-binding peptides were amplified by error-prone polymerase chain reaction with single strand DNA of phages. This peptide contains artificial zinc-binding sequence to restrict peptide conformation. The amplified DNA fragments were digested with Acc65 I and Eag I, and the fragment was ligated into the M13KE phagemid vector. The vector was used to transform host cells (ER2738) by electroporation and the corresponding phage particles were generated. After two cycles of an affinity selection against GM1 monolayer, three positive phage clones were isolated. Phage ELSIA indicated that these clones had affinity for GM1 at 0.01 nM. The binding affinity of an ee2-1 mutant, ee2-15, altered at His-3 to Leu was found to be enhanced. We showed the binding affinity of peptides for target sugar chain was improved by molecular evolution process.

Original languageEnglish
Title of host publicationPolymer Preprints, Japan
Pages1824
Number of pages1
Volume55
Edition1
Publication statusPublished - 2006
Event55th SPSJ Annual Meeting - Nagoya, Japan
Duration: 2006 May 242006 May 26

Other

Other55th SPSJ Annual Meeting
CountryJapan
CityNagoya
Period06/5/2406/5/26

Fingerprint

Scaffolds (biology)
Bacteriophages
Sugars
Peptides
DNA
Polymerase chain reaction
Conformations
Monolayers
Zinc

Keywords

  • Beta structure
  • Ganglioside
  • GM1
  • Molecular evolution
  • Peptide library
  • Phage display method

ASJC Scopus subject areas

  • Engineering(all)

Cite this

Matsubara, T., & Sato, T. (2006). Selection of sugar-binding peptides from a scaffold-containing library. In Polymer Preprints, Japan (1 ed., Vol. 55, pp. 1824)

Selection of sugar-binding peptides from a scaffold-containing library. / Matsubara, Teruhiko; Sato, Toshinori.

Polymer Preprints, Japan. Vol. 55 1. ed. 2006. p. 1824.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Matsubara, T & Sato, T 2006, Selection of sugar-binding peptides from a scaffold-containing library. in Polymer Preprints, Japan. 1 edn, vol. 55, pp. 1824, 55th SPSJ Annual Meeting, Nagoya, Japan, 06/5/24.
Matsubara T, Sato T. Selection of sugar-binding peptides from a scaffold-containing library. In Polymer Preprints, Japan. 1 ed. Vol. 55. 2006. p. 1824
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