Use of a modified bacteriophage to probe the interactions between peptides and ion channel receptors in mammalian cells

Nature Biotechnology
M Li

Abstract

Besides natural peptide ligands, screening of random peptide libraries has yielded novel bioactive peptides for cell surface receptors. A method is described that uses a modified bacteriophage as a detection reagent to monitor the expression of receptor channels in mammalian cells and to probe the molecular interaction between phage-tethered peptides (phi T-peptides) and specific receptor targets. By taking advantage of a specific multivalent interaction between phi T-peptides and the receptor target, assays have been developed that use phi T-peptides specific for the N-methyl-D-aspartate glutamate receptor, an important ligand-gated ion channel in the nervous system, to monitor the receptor expression in cultured mammalian cells. Combining these phi T-peptide binding assays with fluorescence-activated cell sorting, 10(4) random glutamate receptor mutants were screened and candidate interaction residues were identified. This dual heterologous expression system offers a powerful approach to the molecular studies of protein-protein interactions.

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Citations

Feb 14, 2002·Journal of Cellular Biochemistry. Supplement·D Ma, M Li
Oct 11, 2003·Biotechnology Advances·Rita R Colwell
Dec 17, 2009·Proceedings of the National Academy of Sciences of the United States of America·Zoltan TakacsSteve A N Goldstein
Dec 2, 2000·Nature Biotechnology·M Li
Jan 16, 2002·Journal of Receptor and Signal Transduction Research·M Dani
Jan 16, 2002·Journal of Receptor and Signal Transduction Research·M Dani
Apr 23, 2008·Current Protocols in Neuroscience·A Bradbury
Apr 8, 2010·Current Protocols in Neuroscience·Andrew R M Bradbury
Aug 31, 2002·Pediatric and Developmental Pathology : the Official Journal of the Society for Pediatric Pathology and the Paediatric Pathology Society·Felicitas L Lacbawan, Maximilian Muenke

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