Femtosecond X-ray protein nanocrystallography.
Femtosecond X-ray protein nanocrystallography.
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DOI：
10.1038/nature09750
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发表时间：
2011-02-03
期刊：
影响因子：
64.8
通讯作者：
Spence, John C. H.
中科院分区：
文献类型：
--
作者：
Chapman, Henry N.;Fromme, Petra;Barty, Anton;White, Thomas A.;Kirian, Richard A.;Aquila, Andrew;Hunter, Mark S.;Schulz, Joachim;DePonte, Daniel P.;Weierstall, Uwe;Doak, R. Bruce;Maia, Filipe R. N. C.;Martin, Andrew V.;Schlichting, Ilme;Lomb, Lukas;Coppola, Nicola;Shoeman, Robert L.;Epp, Sascha W.;Hartmann, Robert;Rolles, Daniel;Rudenko, Artem;Foucar, Lutz;Kimmel, Nils;Weidenspointner, Georg;Holl, Peter;Liang, Mengning;Barthelmess, Miriam;Caleman, Carl;Boutet, Sebastien;Bogan, Michael J.;Krzywinski, Jacek;Bostedt, Christoph;Bajt, Sasa;Gumprecht, Lars;Rudek, Benedikt;Erk, Benjamin;Schmidt, Carlo;Hoemke, Andre;Reich, Christian;Pietschner, Daniel;Strueder, Lothar;Hauser, Guenter;Gorke, Hubert;Ullrich, Joachim;Herrmann, Sven;Schaller, Gerhard;Schopper, Florian;Soltau, Heike;Kuehnel, Kai-Uwe;Messerschmidt, Marc;Bozek, John D.;Hau-Riege, Stefan P.;Frank, Matthias;Hampton, Christina Y.;Sierra, Raymond G.;Starodub, Dmitri;Williams, Garth J.;Hajdu, Janos;Timneanu, Nicusor;Seibert, M. Marvin;Andreasson, Jakob;Rocker, Andrea;Joensson, Olof;Svenda, Martin;Stern, Stephan;Nass, Karol;Andritschke, Robert;Schroeter, Claus-Dieter;Krasniqi, Faton;Bott, Mario;Schmidt, Kevin E.;Wang, Xiaoyu;Grotjohann, Ingo;Holton, James M.;Barends, Thomas R. M.;Neutze, Richard;Marchesini, Stefano;Fromme, Raimund;Schorb, Sebastian;Rupp, Daniela;Adolph, Marcus;Gorkhover, Tais;Andersson, Inger;Hirsemann, Helmut;Potdevin, Guillaume;Graafsma, Heinz;Nilsson, Bjoern;Spence, John C. H.
x射线晶体学提供了绝大多数的大分子结构，但该方法的成功依赖于生长足够大小的晶体。在传统的测量中，为了记录晶体的数据，必须增加x射线剂量，而太小的晶体在衍射信号被记录之前就会造成大面积的损伤。获得大的、衍射良好的膜蛋白晶体尤其具有挑战性，尽管它们在所有活细胞中都很重要，但只有不到300种独特的结构被确定。在这里，我们提出了一种结构确定的方法，使用来自硬x射线自由电子激光器（直线相干光源）的飞秒脉冲，从完全水化的纳米晶体流中收集单晶x射线衍射“快照”。我们用光系统I的纳米晶体证明了这一概念，光系统I是最大的膜蛋白复合物之一。本研究收集了超过300万张衍射图，并从单个光系统I纳米晶体（~200 nm至2 μm）组装了三维数据集。我们通过使用比大多数损伤过程的时间尺度更短的脉冲来减轻晶体学中的辐射损伤问题。这提供了一种新的方法来确定大分子的结构，这些大分子不能产生足够大小的晶体，用于使用传统辐射源或对辐射损伤特别敏感的研究。
X-ray crystallography provides the vast majority of macromolecular structures, but the success of the method relies on growing crystals of sufficient size. In conventional measurements, the necessary increase in X-ray dose to record data from crystals that are too small leads to extensive damage before a diffraction signal can be recorded. It is particularly challenging to obtain large, well-diffracting crystals of membrane proteins, for which fewer than 300 unique structures have been determined despite their importance in all living cells. Here we present a method for structure determination where single-crystal X-ray diffraction ‘snapshots’ are collected from a fully hydrated stream of nanocrystals using femtosecond pulses from a hard-X-ray free-electron laser, the Linac Coherent Light Source. We prove this concept with nanocrystals of photosystem I, one of the largest membrane protein complexes. More than 3,000,000 diffraction patterns were collected in this study, and a three-dimensional data set was assembled from individual photosystem I nanocrystals (~200 nm to 2 μm in size). We mitigate the problem of radiation damage in crystallography by using pulses briefer than the timescale of most damage processes. This offers a new approach to structure determination of macromolecules that do not yield crystals of sufficient size for studies using conventional radiation sources or are particularly sensitive to radiation damage.
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DOI：
10.1073/pnas.0600973103
发表时间：
2006-03-28
影响因子：
11.1
作者：
Owen, RL;Rudiño-Piñera, E;Garman, EF
通讯作者：
Garman, EF
影响因子：
35
作者：
Emma, P.;Akre, R.;Galayda, J.
通讯作者：
Galayda, J.
影响因子：
3.8
作者：
Kirian RA;Wang X;Weierstall U;Schmidt KE;Spence JC;Hunter M;Fromme P;White T;Chapman HN;Holton J
通讯作者：
Holton J
影响因子：
64.8
作者：
Jordan, P;Fromme, P;Krauss, N
通讯作者：
Krauss, N
影响因子：
19.6
作者：
Fung, Russell;Shneerson, Valentin;Ourmazd, Abbas
通讯作者：
Ourmazd, Abbas