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

Nature Biotechnology volume 15, pages 559–563 (1997)Cite this article

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 (ΦT-peptides) and specific receptor targets. By taking advantage of a specific multivalent interaction between ΦT-peptides and the receptor target, assays have been developed that use ΦT-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 ΦT-peptide binding assays with fluorescence-activated cell sorting, 104 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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Authors and Affiliations

  1. Departments of Physiology and Neuroscience, The Johns Hopkins University School of Medicine, 725 N. Wolfe St., Baltimore, MD, 21205

    Min Li

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Correspondence to Min Li.

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Li, M. Use of a modified bacteriophage to probe the interactions between peptides and ion channel receptors in mammalian cells. Nat Biotechnol 15, 559–563 (1997). https://doi.org/10.1038/nbt0697-559

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  • DOI: https://doi.org/10.1038/nbt0697-559

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