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

Ellmark, Peter LU ; Ohlin, Mats LU ; Borrebaeck, Carl LU and Furebring, Christina LU (2004) In Journal of Molecular Recognition 17(4). p.316-322
Abstract
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... (More)
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. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Molecular Recognition
volume
17
issue
4
pages
316 - 322
publisher
John Wiley & Sons
external identifiers
  • wos:000222513000003
  • scopus:3142603333
ISSN
1099-1352
DOI
10.1002/jmr.678
language
English
LU publication?
yes
id
3d3d15a4-5416-4741-9e86-3d04ad43aa77 (old id 140786)
date added to LUP
2007-06-28 09:36:29
date last changed
2017-01-01 04:30:11
@article{3d3d15a4-5416-4741-9e86-3d04ad43aa77,
  abstract     = {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.},
  author       = {Ellmark, Peter and Ohlin, Mats and Borrebaeck, Carl and Furebring, Christina},
  issn         = {1099-1352},
  language     = {eng},
  number       = {4},
  pages        = {316--322},
  publisher    = {John Wiley & Sons},
  series       = {Journal of Molecular Recognition},
  title        = {A novel mammalian display system for the selection of protein-protein interactions by decoy receptor engagement},
  url          = {http://dx.doi.org/10.1002/jmr.678},
  volume       = {17},
  year         = {2004},
}