Expression of random peptide fused to invasin on bacterial cell surface for selection of cell-targeting peptides

Hiroto Nakajima, Naoki Shimbara, Yoko Shimonishi, Tatsuyuki Mimori, Shin Ichiro Niwa, Hideyuki Saya

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The protein invasin expressed on the cell surface of the pathogenic bacteria Yersinia pseudotuberculosis mediates the entry of this bacterium into cultured mammalian cells. We have developed a system for expression of random peptides on the cell surface of Escherichia coli (E. coli) by creation of a fusion hybrid between a peptide and the invasin protein. The fusion protein constructs consist of part of the outer membrane domain of the invasin protein, six proline spacers, and a decamer of random peptides flanked by cysteine residues (CX10C). Peptides were constitutively expressed on the cell surface in the resulting random decamer peptide library, which we designated as ESPEL (E. coli Surface Peptide Expression Library). The ESPEL was systematically screened for its binding affinity toward human cultured cells. Several bacterial clones were identified whose binding to human cells was mediated by peptides expressed on the bacterial cell surface. Flow cytometric analysis showed that both the identified bacterial clones and these corresponding chemically synthesized peptides bound to human cells specifically. The techniques described provide a new method that uses E. coli random peptide library to select targeting peptides for mammalian cells without any knowledge of the human cellular receptors.

Original languageEnglish
Pages (from-to)121-131
Number of pages11
JournalGene
Volume260
Issue number1-2
DOIs
Publication statusPublished - 2000 Dec 30
Externally publishedYes

Fingerprint

Peptide Library
Peptides
Escherichia coli
Cultured Cells
Clone Cells
Yersinia pseudotuberculosis
Bacteria
Proteins
Proline
Cysteine
Membranes

Keywords

  • Adhesion
  • Escherichia coli
  • Flow cytometric analysis
  • Library

ASJC Scopus subject areas

  • Genetics

Cite this

Expression of random peptide fused to invasin on bacterial cell surface for selection of cell-targeting peptides. / Nakajima, Hiroto; Shimbara, Naoki; Shimonishi, Yoko; Mimori, Tatsuyuki; Niwa, Shin Ichiro; Saya, Hideyuki.

In: Gene, Vol. 260, No. 1-2, 30.12.2000, p. 121-131.

Research output: Contribution to journalArticle

Nakajima, Hiroto ; Shimbara, Naoki ; Shimonishi, Yoko ; Mimori, Tatsuyuki ; Niwa, Shin Ichiro ; Saya, Hideyuki. / Expression of random peptide fused to invasin on bacterial cell surface for selection of cell-targeting peptides. In: Gene. 2000 ; Vol. 260, No. 1-2. pp. 121-131.
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