TY  - JOUR
AU  - Gati, Cornelius
AU  - Oberthuer, Dominik
AU  - Yefanov, Oleksandr
AU  - Bunker
AU  - Stellato, Francesco
AU  - chiu, Elaine
AU  - Yeh, Shin-Mei
AU  - Aquila, Andrew
AU  - Basu, Shibom
AU  - Bean, Richard
AU  - Beyerlein, Kenneth
AU  - Botha, Sabine
AU  - Boutet, Sebastien
AU  - DePonte, Daniel
AU  - Doak, R. Bruce
AU  - Fromme, Raimund
AU  - Galli, Lorenzo
AU  - grotjohan, Ingo
AU  - James, Daniel
AU  - Kupitz, Christopher
AU  - Lomb, Lukas
AU  - Messerschmidt, Marc
AU  - Nass, Karol
AU  - Rendek, Kimberly
AU  - Shoeman, Robert
AU  - Wang, Dingjie
AU  - Weierstall, Uwe
AU  - White, Thomas
AU  - Williams, Garth
AU  - Zatsepin, Nadia
AU  - Fromme, Petra
AU  - Goldie, Kenneth
AU  - Jehle, Johannes
AU  - Metcalf, Peter
AU  - Barty, Anton
AU  - Chapman, Henry N.
TI  - Atomic Structure of Granulin Determined from Native Nanocrystalline Granulovirus Using an X-ray Free-Electron Laser
JO  - Proceedings of the National Academy of Sciences of the United States of America
VL  - 114
IS  - 9
SN  - 0027-8424
CY  - Washington, DC
PB  - National Acad. of Sciences
M1  - PUBDB-2017-00953
SP  - 2247 – 2252
PY  - 2017
N1  - © National Academy of Sciences
AB  - To understand how molecules function in biological systems, new methods are required to obtain atomic resolution structures from biological material under physiological conditions. Intense femtosecond-duration pulses from X-ray free-electron lasers (XFELs) can outrun most damage processes, vastly increasing the tolerable dose before the specimen is destroyed. This in turn allows structure determination from crystals much smaller and more radiation sensitive than previously considered possible, allowing data collection from room temperature structures and avoiding structural changes due to cooling. Regardless, high-resolution structures obtained from XFEL data mostly use crystals far larger than 1 μm<sup>3</sup> in volume, whereas the X-ray beam is often attenuated to protect the detector from damage caused by intense Bragg spots. Here, we describe the 2 Å resolution structure of native nanocrystalline granulovirus occlusion bodies (OBs) that are less than 0.016 μm<sup>3</sup> in volume using the full power of the Linac Coherent Light Source (LCLS) and a dose up to 1.3 GGy per crystal. The crystalline shell of granulovirus OBs consists, on average, of about 9,000 unit cells, representing the smallest protein crystals to yield a high-resolution structure by X-ray crystallography to date. The XFEL structure shows little to no evidence of radiation damage and is more complete than a model determined using synchrotron data from recombinantly produced, much larger, cryocooled granulovirus granulin microcrystals. Our measurements suggest that it should be possible, under ideal experimental conditions, to obtain data from protein crystals with only 100 unit cells in volume using currently available XFELs and suggest that single-molecule imaging of individual biomolecules could almost be within reach.
LB  - PUB:(DE-HGF)16
UR  - <Go to ISI:>//WOS:000395101200053
C6  - pmid:28202732
DO  - DOI:10.1073/pnas.1609243114
UR  - https://bib-pubdb1.desy.de/record/317888
ER  -