Structural enzymology using X-ray free electron lasers.
Structural enzymology using X-ray free electron lasers.
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DOI：
10.1063/1.4972069
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发表时间：
2017-07
期刊：
影响因子：
--
通讯作者：
Schmidt M
中科院分区：
文献类型：
--
作者：
Kupitz C;Olmos JL Jr;Holl M;Tremblay L;Pande K;Pandey S;Oberthür D;Hunter M;Liang M;Aquila A;Tenboer J;Calvey G;Katz A;Chen Y;Wiedorn MO;Knoska J;Meents A;Majriani V;Norwood T;Poudyal I;Grant T;Miller MD;Xu W;Tolstikova A;Morgan A;Metz M;Martin-Garcia JM;Zook JD;Roy-Chowdhury S;Coe J;Nagaratnam N;Meza D;Fromme R;Basu S;Frank M;White T;Barty A;Bajt S;Yefanov O;Chapman HN;Zatsepin N;Nelson G;Weierstall U;Spence J;Schwander P;Pollack L;Fromme P;Ourmazd A;Phillips GN Jr;Schmidt M
混合-注射连续结晶学（MISC）是一种旨在对酶催化反应进行成像的技术，其中小蛋白质晶体在被X射线脉冲探测之前与底物混合。这种方法提供了几个优势超过流动池研究。它提供（i）在近原子分辨率下的室温结构，（ii）范围从微秒到秒的时间分辨率，和（iii）方便的反应引发。它通过使用飞秒X射线脉冲来避免辐射损伤，从而将损伤和化学物质分离开来。在这里，我们证明了MISC是可行的，在X射线自由电子激光通过研究M的反应。结核杆菌β-内酰胺酶微晶体与头孢曲松抗生素溶液。分别在2.8 μ m和2.4 μ m分辨率下获得了载脂蛋白-β-内酰胺酶和头孢曲松结合形式的电子密度图。这些结果为研究循环和非循环反应铺平了道路，并代表了许多底物触发的生物反应的时间分辨结构动力学的新领域。
Mix-and-inject serial crystallography (MISC) is a technique designed to image enzyme catalyzed reactions in which small protein crystals are mixed with a substrate just prior to being probed by an X-ray pulse. This approach offers several advantages over flow cell studies. It provides (i) room temperature structures at near atomic resolution, (ii) time resolution ranging from microseconds to seconds, and (iii) convenient reaction initiation. It outruns radiation damage by using femtosecond X-ray pulses allowing damage and chemistry to be separated. Here, we demonstrate that MISC is feasible at an X-ray free electron laser by studying the reaction of M. tuberculosis ß-lactamase microcrystals with ceftriaxone antibiotic solution. Electron density maps of the apo-ß-lactamase and of the ceftriaxone bound form were obtained at 2.8 Å and 2.4 Å resolution, respectively. These results pave the way to study cyclic and non-cyclic reactions and represent a new field of time-resolved structural dynamics for numerous substrate-triggered biological reactions.
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DOI：
10.1126/science.1217737
发表时间：
2012-07-20
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Boutet S;Lomb L;Williams GJ;Barends TR;Aquila A;Doak RB;Weierstall U;DePonte DP;Steinbrener J;Shoeman RL;Messerschmidt M;Barty A;White TA;Kassemeyer S;Kirian RA;Seibert MM;Montanez PA;Kenney C;Herbst R;Hart P;Pines J;Haller G;Gruner SM;Philipp HT;Tate MW;Hromalik M;Koerner LJ;van Bakel N;Morse J;Ghonsalves W;Arnlund D;Bogan MJ;Caleman C;Fromme R;Hampton CY;Hunter MS;Johansson LC;Katona G;Kupitz C;Liang M;Martin AV;Nass K;Redecke L;Stellato F;Timneanu N;Wang D;Zatsepin NA;Schafer D;Defever J;Neutze R;Fromme P;Spence JC;Chapman HN;Schlichting I
通讯作者：
Schlichting I
影响因子：
56.9
作者：
Schotte, F;Lim, MH;Anfinrud, PA
通讯作者：
Anfinrud, PA
影响因子：
21.8
作者：
通讯作者：
--
影响因子：
3.7
作者：
Feiler, Christian;Fisher, Adam C.;DeLisa, Matthew P.
通讯作者：
DeLisa, Matthew P.
影响因子：
5.7
作者：
Geremia, S;Campagnolo, M;Johnson, LN
通讯作者：
Johnson, LN