Journal article

Direct Molecular Evolution of Detergent-Stable G Protein-Coupled Receptors Using Polymer Encapsulated Cells

Daniel J Scott, Andreas Plueckthun

JOURNAL OF MOLECULAR BIOLOGY | ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD | Published : 2013

Abstract

G protein-coupled receptors (GPCRs) are the largest class of pharmaceutical protein targets, yet drug development is encumbered by a lack of information about their molecular structure and conformational dynamics. Most mechanistic and structural studies as well as in vitro drug screening with purified receptors require detergent solubilization of the GPCR, but typically, these proteins exhibit only low stability in detergent micelles. We have developed the first directed evolution method that allows the direct selection of GPCRs stable in a chosen detergent from libraries containing over 100 million individual variants. The crucial concept was to encapsulate single Escherichia coli cells of ..

View full abstract

Grants

Funding Acknowledgements

We acknowledge the professional help given by the staff of the Center for Microscopy and Image Analysis at the University of Zurich, where all FACS and microscopy were performed, particularly Vinko Tosevski, Claudia Dumrese and Andres Kaech. This work was supported by a C. J. Martin Biomedical Overseas Fellowship from the National Health and Medical Research Council of Australia. and a postdoctoral fellowship from the Forschungskredit of the University of Zurich (both to D.J.S.) and by the Swiss National Center of Competence in Research Structural Biology (to A.P.).