Viscosity-adjustable grease matrices for serial nanocrystallography
Viscosity-adjustable grease matrices for serial nanocrystallography
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用于连续纳米晶体学的粘度可调润滑脂基质
DOI：
10.1038/s41598-020-57675-7
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发表时间：
2020
影响因子：
4.6
通讯作者：
Ishikawa Tetsuya
中科院分区：
文献类型：
--
作者：
Sugahara Michihiro;Motomura Koji;Suzuki Mamoru;Masuda Tetsuya;Joti Yasumasa;Numata Keiji;Tono Kensuke;Yabashi Makina;Ishikawa Tetsuya
连续飞秒晶体学（SFX）能够以最小的辐射损伤测定蛋白质的室温结构。在x射线自由电子激光器的光束路径中引入了一种高粘性的润滑脂基质作为晶体载流子，以~0.5 μl min - 1的低流速连续加载样品。该基质使测定蛋白质结构的样品消耗量小于1毫克的蛋白质成为可能。在高粘度微挤压进样器中，基质的粘度是保证样品柱连续稳定的重要因素。使用传统的商业润滑脂（一种油基的粘性剂），如果对基质的粘度控制不足，通常会产生意想不到的低粘度，从而提供不稳定的样品流，并产生诸如流卷曲等效应。润滑脂粘度的调整是极其困难的，因为商业润滑脂含有未知的化合物，这可能是意想不到的蛋白质抑制剂。本研究引入了两种新型润滑脂基质载体，其中包含比传统润滑脂粘度更高的已知化合物，以确定纳米/微晶体的蛋白酶K结构。
Serial femtosecond crystallography (SFX) has enabled determination of room temperature structures of proteins with minimum radiation damage. A highly viscous grease matrix acting as a crystal carrier for serial sample loading at a low flow rate of ~0.5 μl min−1was introduced into the beam path of X-ray free-electron laser. This matrix makes it possible to determine the protein structure with a sample consumption of less than 1 mg of the protein. The viscosity of the matrix is an important factor in maintaining a continuous and stable sample column from a nozzle of a high viscosity micro-extrusion injector for serial sample loading. Using conventional commercial grease (an oil-based, viscous agent) with insufficient control of viscosity in a matrix often gives an unexpectedly low viscosity, providing an unstable sample stream, with effects such as curling of the stream. Adjustment of the grease viscosity is extremely difficult since the commercial grease contains unknown compounds, which may act as unexpected inhibitors of proteins. This study introduces two novel grease matrix carriers comprising known compounds with a viscosity higher than that of conventional greases, to determine the proteinase K structure from nano-/microcrystals.
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影响因子：
3.9
作者：
Kovácsová G;Grünbein ML;Kloos M;Barends TRM;Schlesinger R;Heberle J;Kabsch W;Shoeman RL;Doak RB;Schlichting I
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Liu W
DOI：
10.1126/science.1244142
发表时间：
2013-12-20
期刊：
Science (New York, N.Y.)
影响因子：
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作者：
Liu W;Wacker D;Gati C;Han GW;James D;Wang D;Nelson G;Weierstall U;Katritch V;Barty A;Zatsepin NA;Li D;Messerschmidt M;Boutet S;Williams GJ;Koglin JE;Seibert MM;Wang C;Shah ST;Basu S;Fromme R;Kupitz C;Rendek KN;Grotjohann I;Fromme P;Kirian RA;Beyerlein KR;White TA;Chapman HN;Caffrey M;Spence JC;Stevens RC;Cherezov V
通讯作者：
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影响因子：
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作者：
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通讯作者：
T. Nakane
影响因子：
64.8
作者：
Kang Y;Zhou XE;Gao X;He Y;Liu W;Ishchenko A;Barty A;White TA;Yefanov O;Han GW;Xu Q;de Waal PW;Ke J;Tan MH;Zhang C;Moeller A;West GM;Pascal BD;Van Eps N;Caro LN;Vishnivetskiy SA;Lee RJ;Suino-Powell KM;Gu X;Pal K;Ma J;Zhi X;Boutet S;Williams GJ;Messerschmidt M;Gati C;Zatsepin NA;Wang D;James D;Basu S;Roy-Chowdhury S;Conrad CE;Coe J;Liu H;Lisova S;Kupitz C;Grotjohann I;Fromme R;Jiang Y;Tan M;Yang H;Li J;Wang M;Zheng Z;Li D;Howe N;Zhao Y;Standfuss J;Diederichs K;Dong Y;Potter CS;Carragher B;Caffrey M;Jiang H;Chapman HN;Spence JC;Fromme P;Weierstall U;Ernst OP;Katritch V;Gurevich VV;Griffin PR;Hubbell WL;Stevens RC;Cherezov V;Melcher K;Xu HE
通讯作者：
Xu HE