000000687 001__ 687
000000687 005__ 20190130104941.0
000000687 02470 $$a10.1073/pnas.1609243114$$2doi
000000687 037__ $$aPROCEEDING-2017-004
000000687 041__ $$aeng
000000687 245__ $$aAtomic structure of granulin determined from native nanocrystalline granulovirus using an X-ray free-electron laser
000000687 260__ $$aWashington, DC$$bNational Acad. of Sciences$$c2017$$hUSA
000000687 269__ $$a2017
000000687 336__ $$aProceedings
000000687 520__ $$aTo 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 $μm^{3}$ 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 $μm^{3}$ 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.
000000687 546__ $$aEnglish
000000687 6531_ $$aXFEL
000000687 6531_ $$ananocrystals
000000687 6531_ $$astructural biology
000000687 6531_ $$abioimaging
000000687 6531_ $$aSFX
000000687 655__ $$aOther
000000687 690__ $$aExternal experiment
000000687 7001_ $$aGati, Cornelius
000000687 7001_ $$aOberthuer, Dominik
000000687 7001_ $$aYefanov, Oleksandr
000000687 7001_ $$aBunker, Richard D.
000000687 7001_ $$aStellato, Francesco
000000687 7001_ $$aChiu, Elaine
000000687 7001_ $$aYeh, Shin-Mei
000000687 7001_ $$aAquila, Andrew
000000687 7001_ $$aBasu, Shibom
000000687 7001_ $$01257$$aBean, Richard
000000687 7001_ $$aBeyerlein, Kenneth R.
000000687 7001_ $$aBotha, Sabine
000000687 7001_ $$aBoutet, Sébastien
000000687 7001_ $$aDePonte, Daniel P.
000000687 7001_ $$aDoak, R. Bruce
000000687 7001_ $$aFromme, Raimund
000000687 7001_ $$aGalli, Lorenzo
000000687 7001_ $$aGrotjohann, Ingo
000000687 7001_ $$aJames, Daniel R.
000000687 7001_ $$aKupitz, Christopher
000000687 7001_ $$aLomb, Lukas
000000687 7001_ $$01252$$aMesserschmidt, Marc
000000687 7001_ $$aNass, Karol
000000687 7001_ $$aRendek, Kimberly
000000687 7001_ $$aShoeman, Robert L.
000000687 7001_ $$aWang, Dingjie
000000687 7001_ $$aWeierstall, Uwe
000000687 7001_ $$aWhite, Thomas A.
000000687 7001_ $$aWilliams, Garth J.
000000687 7001_ $$aZatsepin, Nadia A.
000000687 7001_ $$aFromme, Petra
000000687 7001_ $$aSpence, John C.H.
000000687 7001_ $$aGoldie, Kenneth N.
000000687 7001_ $$aJehle, Johannes A.
000000687 7001_ $$aMetcalf, Peter
000000687 7001_ $$aBarty, Anton
000000687 7001_ $$aChapman, Henry N.
000000687 773__ $$n9$$q2247-2252$$tProceedings of PNAS 2017$$v114
000000687 790__ $$aEuXFEL staff
000000687 790__ $$aOther
000000687 85641 $$uhttp://www.pnas.org/content/early/2017/02/14/1609243114
000000687 8560_ $$fkurt.ament@xfel.eu
000000687 900__ $$aInstrument SPB/SFX
000000687 980__ $$aPROCEEDING