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In-house UV radiation-damage-induced phasing of selenomethionine-labeled protein structures |
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| Authors: | Pedro José Barbosa Pereira Antoine Royant Santosh Panjikar Daniele de Sanctis |
| Institution: | 1. IBMC – Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua do Campo Alegre 823, 4150-180 Porto, Portugal;2. Institut de Biologie Structurale Jean-Pierre Ebel, UMR 5075 CNRS, Commissariat à l’Energie Atomique, Université Joseph Fourier, 41 rue Jules Horowitz, 38027 Grenoble Cedex, France;3. Structural Biology Group, European Synchrotron Radiation Facility, 6 rue Jules Horowitz, 38043 Grenoble Cedex, France;4. Australian Synchrotron, 800 Blackburn Road, Clayton, Victoria 3168, Australia;5. Department of Biochemistry and Molecular Biology, Monash University, Victoria 3800, Australia |
| Abstract: | Selenomethionine labeling is the most common technique used in protein crystallography to derivatize recombinant proteins for experimental phasing using anomalous scattering at tunable synchrotron beamlines. Recently, it has been shown that UV radiation depletes electron density of selenium atoms of selenomethionine residues and that UV radiation-damage-induced phasing (equivalent to single isomorphous replacement) protocol can be applied to calculate experimental phases. Here we present the straightforward integration of a UV source with an in-house diffractometer. We show how this setup can extend the capabilities of a sealed tube X-ray generator and be used for experimental phasing of selenium-labeled proteins. |
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