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Avidity-based Extracellular Interaction Screening (AVEXIS) for the Scalable Detection of Low-affinity Extracellular Receptor-Ligand Interactions
Published on: March 5, 2012
Cell growth selection system to detect extracellular and transmembrane protein interactions
David M Urech1, Peter Lichtlen, Alcide Barberis
1ESBATech AG, Wagistrasse 21, CH-8952 Zürich-Schlieren, Switzerland. urech@esbatech.com
Biochimica Et Biophysica Acta
|July 26, 2003
Summary
This study introduces SCINEX-P, a novel yeast-based system for detecting extracellular protein interactions. It overcomes limitations of traditional methods by assaying interactions in an oxidizing environment, crucial for protein folding.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Extracellular and cell surface protein interactions are vital in physiological and pathological processes.
- Disulfide bond formation in the oxidizing secretory pathway is critical for protein folding and stability.
- Conventional yeast two-hybrid assays occur in a reducing environment, limiting their ability to detect extracellular interactions.
Purpose of the Study:
- To develop a novel system for detecting extracellular and membrane protein interactions.
- To overcome the limitations of existing methods in studying extracellular protein interactions.
Main Methods:
- Exploited yeast Ire1p, an endoplasmic reticulum (ER) membrane protein involved in the unfolded protein response (UPR).
- Developed SCINEX-P (screening for interactions between extracellular proteins) system.
- Fused proteins of interest to truncated Ire1p, replacing its N-terminal lumenal domain (NLD).
Main Results:
- Specific interactions between protein partners induced dimerization of the Ire1p moiety.
- Dimerization activated the endogenous UPR signaling pathway.
- Activation led to transcription of genes enabling cell growth under selective conditions.
Conclusions:
- SCINEX-P effectively detects extracellular protein-protein interactions.
- The system leverages the UPR pathway for signal transduction.
- SCINEX-P provides a valuable tool for studying extracellular and membrane protein interactions in a relevant cellular context.
