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2 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.
iCLIP - Transcriptome-wide Mapping of Protein-RNA Interactions with Individual Nucleotide Resolution Julian Konig1, Kathi Zarnack2, Gregor Rot3, Tomaz Curk3, Melis Kayikci1, Blaz Zupan3, Daniel J. Turner4, Nicholas M. Luscombe2, Jernej Ule1 1Laboratory of Molecular Biology, Medical Research Council - MRC, 2European Bioinformatics Institute, EMBL Heidelberg, 3Computer and Information Science, University of Ljubljana, 4Wellcome Trust Genome Campus, Wellcome Trust Sanger Institute The spatial arrangement of RNA-binding proteins on a transcript is a key determinant of post-transcriptional regulation. Therefore, we developed individual-nucleotide resolution UV crosslinking and immunoprecipitation (iCLIP) that allows precise genome-wide mapping of the binding sites of an RNA-binding protein.