Abstract
Protein crystallization in cells has been observed several times in nature. However, owing to their small size these crystals have not yet been used for X-ray crystallographic analysis. We prepared nano-sized in vivo-grown crystals of Trypanosoma brucei enzymes and applied the emerging method of free-electron laser-based serial femtosecond crystallography to record interpretable diffraction data. This combined approach will open new opportunities in structural systems biology.
| Original language | English |
|---|---|
| Journal | Nature Methods |
| Volume | 9 |
| Issue number | 3 |
| Pages (from-to) | 259-262 |
| Number of pages | 4 |
| ISSN | 1548-7091 |
| DOIs | |
| Publication status | Published - 01.03.2012 |
Funding
FEL experiments were carried out at LCLS in June 2010 (TbCatB) and in August 2011 (TbIMPDH), a national user facility operated by Stanford University on behalf of the US Department of Energy, Office of Basic Energy Sciences. The X-ray diffraction experiments on recrystallized TbCatB crystals were carried out at beamline X06DA of the Swiss Light Source (Villigen, Switzerland). This work was supported in part by a grant from the Deutsche Forschungsgemeinschaft (DFG), from the Swedish Research Council, from the Knut och Alice Wallenbergs Stiftelse, from the European Research Council, as well as by US National Science Foundation award MCB-1021557. R.K. received a fellowship from the Landesgraduiertenförderung Baden-Württemberg. L.R., D. Rehders and C. Betzel thank the German Federal Ministry for Education and Research for funding (grants 01KX0806 and 01KX0807). Support from the Hamburg Ministry of Science and Research and Joachim Herz Stiftung as part of the Hamburg Initiative for Excellence in Research and the Hamburg School for Structure and Dynamics in infection, and from the DFG Cluster of Excellence “Inflammation at Interfaces” (EXC 306) is gratefully acknowledged. Funding for the development and operation of the CFEL-ASG multipurpose (CAMP) instrument within the Advanced Study Group at the Center for Free-Electron Laser Science was provided by the Max Planck Society. M.J.B., R.G.S. and C.Y.H. acknowledge funding from the US Department of Energy Office of Basic Energy Sciences through the Photon Ultrafast Laser Science and Engineering (PULSE) Institute at the Stanford Linear Accelerator Center (SLAC) National Accelerator Laboratory. 1Interfaculty Institute of Biochemistry, University of Tübingen, Tübingen, Germany.2Joint Laboratory for Structural Biology of Infection and Inflammation, Institute of Biochemistry and Molecular Biology, University of Hamburg, and Institute of Biochemistry, University of Lübeck, at Deutsches Elektronen-Synchrotron (DESY), Hamburg, Germany. 3Institute for Experimental Physics, University of Hamburg, Hamburg, Germany. 4Center for Free-Electron Laser Science, DESY, Hamburg, Germany. 5Laboratory of Molecular Biophysics, Department of Cell and Molecular Biology, Uppsala University, Uppsala, Sweden.6Photon Science, DESY, Hamburg, Germany. 7Photon Ultrafast Laser Science and Engineering (PULSE) Institute, Stanford Linear Accelerator Center (SLAC) National Accelerator Laboratory, Menlo Park, California, USA. 8Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California, USA.9Department of Physics, Arizona State University, Tempe, Arizona, USA. 10Max Planck Advanced Study Group, Center for Free-Electron Laser Science, Hamburg, Germany.11Max-Planck-Institut für Kernphysik, Heidelberg, Germany. 12Max-Planck-Institut für Medizinische Forschung, Heidelberg, Germany. 13PNSensor GmbH, Munich, Germany. 14Max-Planck-Institut Halbleiterlabor, Munich, Germany. 15Department of Chemistry and Biochemistry, Arizona State University, Tempe, Arizona, USA.16National Energy Research Scientific Computing Center, Lawrence Berkeley National Laboratory, Berkeley, California, USA.17Max-Planck-Institut für extraterrestrische Physik, Garching, Germany. 18University of Siegen, Siegen, Germany. 19Department of Pediatrics, Vanderbilt University School of Medicine, Nashville, Tennessee, USA.20Institute of Biochemistry and Molecular Biology, University of Hamburg, Hamburg, Germany.21Present addresses: European XFEL GmbH, Hamburg, Germany (A.A. and N.C.) and Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California, USA (J.H.).22These authors contributed equally to this work. Correspondence should be addressed to M.D. ([email protected]).
