3 articles published in JoVE
Strategic Screening and Characterization of the Visual GPCR-mini-G Protein Signaling Complex for Successful Crystallization Filip Pamula1,2, Jonas Mühle1, Alain Blanc3, Rony Nehmé4, Patricia C. Edwards4, Christopher G. Tate4, Ching-Ju Tsai1 1Laboratory of Biomolecular Research, Paul Scherrer Institute, 2Department of Biology, ETH Zürich, 3Center for Radiopharmaceutical Sciences, Paul Scherrer Institute, 4Laboratory of Molecular Biology, Medical Research Council This report describes screening of different detergents for preparing the visual GPCR, rhodopsin, and its complex with mini-Go. Biochemical methods characterizing the quality of the complex at different stages during purification are demonstrated. This protocol can be generalized to other membrane protein complexes for their future structural studies.
Improving High Viscosity Extrusion of Microcrystals for Time-resolved Serial Femtosecond Crystallography at X-ray Lasers Daniel James1, Tobias Weinert1, Petr Skopintsev1, Antonia Furrer1, Dardan Gashi1,2, Tomoyuki Tanaka3,4, Eriko Nango3,4, Przemyslaw Nogly1,5, Joerg Standfuss1 1Division of Biology and Chemistry - Laboratory for Biomolecular Research, Paul Scherrer Institut, 2Photon Science Division - SwissFEL, Paul Scherrer Institut, 3RIKEN SPring-8 Center, 4Department of Cell Biology, Graduate School of Medicine, Kyoto University, 5Department of Biology, ETH Zürich The success of a time-resolved serial femtosecond crystallography experiment is dependent on efficient sample delivery. Here, we describe protocols to optimize the extrusion of bacteriorhodopsin microcrystals from a high viscosity micro-extrusion injector. The methodology relies on sample homogenization with a novel three-way coupler and visualization with a high-speed camera.
A Practical Guide on Coupling a Scanning Mobility Sizer and Inductively Coupled Plasma Mass Spectrometer (SMPS-ICPMS) Mohamed Tarik1, Debora Foppiano1,2, Adrian Hess1,3, Christian Ludwig1,2 1Bioenergy and Catalysis Laboratory (LBK), Energy and Environment Research Division (ENE), Paul Scherrer Institute, 2Environmental Engineering Institute (IIE), School of Architecture, Civil and Environmental Engineering (ENAC), École; Polytechnique Fédérale de Lausanne (EPFL), 3Institute for Atmospheric and Climate Science ETH Zurich In this work a practical guide is provided, describing the different steps to establish the coupling of SMPS and ICPMS systems, and how to use them. Three descriptive examples are presented.