Screening and characterization of affinity peptide tags specific to polystyrene supports for the orientated immobilization of proteins

Yoichi Kumada, Yasuhide Tokunaga, Hiroyuki Imanaka, Koreyoshi Imamura, Takaharu Sakiyama, Shigeo Katoh, Kazuhiro Nakanishi

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

Dodecapeptides that exhibit a high affinity specific to a polystyrene surface (PS-tags) were screened using an Escherichia coli random peptide display library system, and the compounds were used as a peptide tag for the site-specific immobilization of proteins. The various PS-tags obtained after 10 rounds of biopanning selection were mainly composed of basic and aliphatic amino acid residues, most of which were arranged in close proximity to one another. Mutant-type glutathione S-transferases (GSTs) fused with the selected PS-tags, PS19 (RAFIASRRIKRP) and PS23 (AGLRLKKAAIHR) at their C-terminus, GST-PS19 and GST-PS23, when adsorbed on the PS latex beads had a higher affinity than the wild-type GST, and the specific remaining activity of the immobilized mutant-type GSTs was approximately 10 times higher than that of the wild-type GST. The signal intensity detected for GST-PS19 and GST-PS23 adsorbed on hydrophilic and hydrophobic PS surfaces using an anti-peptide antibody specific for the N-terminus peptide of GST was much higher than that for the wild-type GST. These findings indicate that the mutant-type GSTs fused with the selected peptide tags, PS19 and PS23, could be site-specifically immobilized on the surface of polystyrene with their N-terminal regions directed toward the solution. Thus, the selected peptide tags would be useful for protein immobilization in the construction of enzyme-linked immunosorbent assay (ELISA) systems and protein-based biochips.

Original languageEnglish
Pages (from-to)401-405
Number of pages5
JournalBiotechnology Progress
Volume22
Issue number2
DOIs
Publication statusPublished - Mar 1 2006

ASJC Scopus subject areas

  • Biotechnology

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