Du är här

A novel mammalian display system for the selection of protein-protein interactions by decoy receptor engagement

Publiceringsår: 2004
Språk: Engelska
Sidor: 316-322
Publikation/Tidskrift/Serie: Journal of Molecular Recognition
Volym: 17
Nummer: 4
Dokumenttyp: Artikel
Förlag: John Wiley & Sons, Ltd

Sammanfattning

The emerging field of proteomics has created a need for new high-throughput methodologies for the analysis of gene products. An attractive approach is to develop systems that allow for clonal selection of interacting protein pairs from large molecular libraries. In this study, we have characterized a novel approach for identification and selection of protein-protein interactions, denoted SPIRE (selection of protein interactions by receptor engagement), which is based on a mammalian expression system. We have demonstrated proof of concept by creating a general plasma membrane bound decoy receptor, by displaying a protein or a peptide genetically fused to a trunctated version of the CD40 molecule. When this decoy receptor is engaged by a ligand to the displayed protein/peptide, the receptor expressing cell is rescued from apoptosis. To design a high-throughput system with a highly parallel capacity, we utilized the B cell line WEHI-231, as carrier of the decoy receptor. One specific peptide-displaying cell could be identified and amplified, based on a specific receptor engagement, in a background of 12 500 wild-type cells after four selections. This demonstrates that the approach may serve as a tool in post-genomic research for identifying protein-protein interactions, without prior knowledge of either component. Copyright (C) 2004 John Wiley Sons, Ltd.

Disputation

Nyckelord

  • Medicine and Health Sciences
  • Chemistry

Övrigt

Published
Yes
  • ISSN: 0952-3499

Box 117, 221 00 LUND
Telefon 046-222 00 00 (växel)
Telefax 046-222 47 20
lu [at] lu [dot] se

Fakturaadress: Box 188, 221 00 LUND
Organisationsnummer: 202100-3211
Om webbplatsen

LERU logo U21 logo