Watching a protein as it functions with 150-ps time-resolved X-ray crystallography
Watching a protein as it functions with 150-ps time-resolved X-ray crystallography
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DOI：
10.1126/science.1078797
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发表时间：
2003-06-20
期刊：
影响因子：
56.9
通讯作者：
Anfinrud, PA
中科院分区：
文献类型：
--
作者：
Schotte, F;Lim, MH;Anfinrud, PA
我们报告皮秒时间分辨x射线衍射从肌红蛋白（Mb）突变体，其中Leu（29）被Phe （L29F突变体）取代。每一帧的结构演变，分辨率为1.8埃，允许人们真正“观察”蛋白质执行其功能。对闪光降解的l29fmbco进行了时间分辨中红外光谱分析，发现了一种短寿命的CO中间体，其140-ps的寿命比野生型蛋白短1000倍。蛋白质的电子密度图揭示了比羧基和脱氧状态之间的结构差异更剧烈的瞬态构象变化，并描绘了相关的侧链运动，负责迅速将CO从其主要对接位点扫除。
We report picosecond time-resolved x-ray diffraction from the myoglobin (Mb) mutant in which Leu(29) is replaced by Phe (L29F mutant). The frame-by-frame structural evolution, resolved to 1.8 angstroms, allows one to literally "watch" the protein as it executes its function. Time-resolved mid-infrared spectroscopy of flash-photolyzed L29F MbCO revealed a short-lived CO intermediate whose 140-ps lifetime is shorter than that found in wild-type protein by a factor of 1000. The electron density maps of the protein unveil transient conformational changes far more dramatic than the structural differences between the carboxy and deoxy states and depict the correlated side-chain motion responsible for rapidly sweeping CO away from its primary docking site.