High-speed fixed-target serial virus crystallography

被引:0
|
作者
Roedig, Philip [1 ]
Ginn, Helen M. [2 ,3 ]
Pakendorf, Tim [1 ]
Sutton, Geoff [2 ]
Harlos, Karl [2 ]
Walter, Thomas S. [2 ]
Meyer, Jan [1 ]
Fischer, Pontus [1 ]
Duman, Ramona [3 ]
Vartiainen, Ismo [4 ]
Reime, Bernd [1 ]
Warmer, Martin [1 ]
Brewster, Aaron S. [5 ]
Young, Iris D. [5 ]
Michels-Clark, Tara [5 ]
Sauter, Nicholas K. [5 ]
Kotecha, Abhay [2 ]
Kelly, James [6 ,7 ]
Rowlands, David J. [6 ]
Sikorsky, Marcin [8 ]
Nelson, Silke [8 ]
Damiani, Daniel S. [8 ]
Alonso-Mori, Roberto [8 ]
Ren, Jingshan [2 ]
Fry, Elizabeth E. [2 ]
David, Christian [9 ]
Stuart, David I. [2 ,3 ]
Wagner, Armin [3 ]
Meents, Alke [1 ,10 ]
机构
[1] Deutsch Elekt Synchrotron DESY, Photon Sci, Hamburg, Germany
[2] Univ Oxford, Wellcome Trust Ctr Human Genet, Div Struct Biol, Oxford, England
[3] Diamond Light Source Ltd, Didcot, Oxon, England
[4] Univ Eastern Finland, Inst Photon, Joensuu, Finland
[5] Lawrence Berkeley Natl Lab, Mol Biophys & Integrated Bioimaging Div, Berkeley, CA USA
[6] Univ Leeds, Fac Biol Sci, Sch Mol & Cellular Biol, Leeds, W Yorkshire, England
[7] Pirbright Inst, Pirbright, England
[8] SLAC Natl Accelerator Lab, Linac Coherent Light Source, Menlo Pk, CA USA
[9] Paul Scherrer Inst, Villigen, Switzerland
[10] Ctr Free Elect Laser Sci CFEL, Hamburg, Germany
基金
美国国家卫生研究院; 英国医学研究理事会; 英国惠康基金;
关键词
PATTERNED SILICON CHIP; FEMTOSECOND CRYSTALLOGRAPHY; DIFFRACTION; PROTEINS; HUMIDITY; ENTRY;
D O I
10.1038/NMETH.4335
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We report a method for serial X-ray crystallography at X-ray free-electron lasers (XFELs), which allows for full use of the current 120-Hz repetition rate of the Linear Coherent Light Source (LCLS). Using a micropatterned silicon chip in combination with the high-speed Roadrunner goniometer for sample delivery, we were able to determine the crystal structures of the picornavirus bovine enterovirus 2 (BEV2) and the cytoplasmic polyhedrosis virus type 18 polyhedrin, with total data collection times of less than 14 and 10 min, respectively. Our method requires only micrograms of sample and should therefore broaden the applicability of serial femtosecond crystallography to challenging projects for which only limited sample amounts are available. By synchronizing the sample exchange to the XFEL repetition rate, our method allows for most efficient use of the limited beam time available at XFELs and should enable a substantial increase in sample throughput at these facilities.
引用
收藏
页码:805 / +
页数:9
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