Conformational Thermo-stabilization of Functional Biomolecular Modules and development of light-con-trolled Molecular Switches

Conformational Thermo-stabilization of Functional Biomolecular Modules and development of light-con-trolled Molecular Switches

  • Simplified scheme of GPCR signalling: Light or ligand binding triggers a conformational change of the receptor that induces G protein activation.
    Simplified scheme of GPCR signalling: Light or ligand binding triggers a conformational change of the receptor that induces G protein activation.

The overarching aim of this project is to engineer and produce stable GPCRs, GPCR complexes, G proteins, and arrestins and explore their potential for applications in complex molecular systems.

Future G protein-coupled receptors (GPCR) platforms will require the availability of stable, robust, cell-free signalling complexes comprising receptor and effector proteins. An expert in the field of GPCRs, project leader Gerhard Schertler has determined the first three-dimensional structure of a GPCR by electron crystallography. This structure of the light receptor rhodopsin provided the first experimental evidence for the arrangement of the seven trans-membrane helices of the receptor within the membrane and, therefore, served as a first experimental template for modelling the structures of other class A GPCRs.

GPCRs are integral membrane proteins that play a fundamental role in many physiological and pathological processes of the visual, endocrine, olfactory and nervous systems. Class A GPCRs represent the largest protein family in the human genome with almost 1000 members. Although GPCRs are the target of ~30% of all medical drugs, their potential remains largely untapped. This impressively illustrates their huge medical importance.

The functionality of candidate receptor variants for potential nano- and bio-technological applications will be tested in a cell-free environment by reconstituting them into lipid bilayers and block co-polymers. The PSI protein production platform for the expression of recombinant proteins with a particular focus of GPCRs, arrestins and G proteins has high-throughput capabilities for cloning and protein purification.

Articles

M. Hilbert, A. Noga, D. Frey, V. Hamel, P. Guichard, S. H. Kraatz, M. Pfreundschuh, S. Hosner, I. Flückiger, R. Jaussi, M. M. Wieser, K. M. Thieltges, X. Deupi, D. J. Mülle, R. Kammerer, P. Gönczy, M. Hirono, M. O. Steinmetz, D. J. MüllerSAS-6 engineering reveals interdependence between cartwheel and microtubules in determining centriole architecture“ Nat. Cell Biol. 18, 393 (2016). [Link]
D. Sun, T. Flock, X. Deupi, S. Maeda, M. Matkovic, S. Mendieta, D. Mayer, R. J. Dawson, G. Schertler, M. M. Babu, D. B. Veprintsev “Probing Gαi1 protein activation at single–amino acid resolution“ Nat. Struct. Mol. Biol. 22, 686 (2015). [Link]
F. M. Heydenreich, Z. Vuckovic, M. Matkovic, D. B. Veprintsev “Stabilization of G protein-coupled receptors by point mutations“ Front. Pharmacol. 6, 82 (2015). [Link] [More Information]
D. Milić, D. B. Veprintsev “Large-scale production and protein engineering of G protein-coupled receptors for structural studies“ Front. Pharmacol. 6, 66 (2015). [Link] [More Information]

Who works with whom?

Prof. Gebhard F.X. Schertler and Dr. Richard A. Kammerer from the Laboratory of Biomolecular Research of Paul Scherrer Institute (PSI) lead this project together. They work with PhD student Martin Spillmann.

The spin-off company InterAx evolved out of the NCCR-project at PSI.

Group

Read more about the Schertler/Kammerer-Group here.

Collaborations

Their experiments will be carried out in close collaboration with projects lead by Dimitrios Fotiadis, Wolfgang Meier and Cornelia Palivan, Yaakov Benenson, Martin Fussenegger, Sven Panke and Botond Roska.