Fourme R, Kahn R, Mezouar M, Girard E, Hoerentrup C, Prangé T, Ascone I
LURE, UMR 130 CNRS-CEA-MRT, Bâtiment 209D, Université Paris-Sud, F91898 Orsay CEDEX, France.
J Synchrotron Radiat. 2001 Sep 1;8(Pt 5):1149-56. doi: 10.1107/s0909049501011037. Epub 2001 Aug 28.
A new set-up and associated methodology for the collection of angle-dispersive diffraction data from protein crystals submitted to high hydrostastic pressure have been developed on beamline ID30 at the ESRF. The instrument makes use of intense X-rays of ultra-short wavelength emitted by two collinear undulators, and combines a membrane-driven diamond-anvil cell mounted on a two-axis goniometer and an imaging-plate scanner. Sharp and clean diffraction pictures from tetragonal crystals of hen egg-white lysozyme (tHEWL) and orthorhombic crystals of bovine erythrocyte Cu, Zn superoxide dismutase (SOD) were recorded at room temperature and pressures up to 0.915 and 1.00 GPa, respectively. The compressibility of tHEWL was determined from unit-cell parameters determined at 24 different pressures up to 0.915 GPa. High-pressure diffraction data sets from several crystals of tHEWL were collected and analyzed. Merging of data recorded on different crystals at 0.30 and 0.58 GPa produced two sets of structure amplitudes with good resolution, completeness, redundancy and R(sym) values. A third set at 0.69 GPa was of a similar quality except a lower completeness. The three structures have been refined. The pressure-induced loss of crystalline order in a tHEWL crystal beyond 0.82 GPa was captured through a series of diffraction pictures.
在欧洲同步辐射装置（ESRF）的ID30光束线上，已开发出一种新装置及相关方法，用于收集承受高静水压力的蛋白质晶体的角散射衍射数据。该仪器利用两个共线波荡器发出的超短波长强X射线，并结合安装在双轴测角仪上的膜驱动金刚石对顶砧池和成像板扫描仪。在室温以及分别高达0.915 GPa和1.00 GPa的压力下，记录到了来自蛋清溶菌酶（tHEWL）四方晶体和牛红细胞铜锌超氧化物歧化酶（SOD）正交晶体的清晰锐利的衍射图像。tHEWL的压缩性由在高达0.915 GPa的24个不同压力下测定的晶胞参数确定。收集并分析了来自多个tHEWL晶体的高压衍射数据集。在0.30 GPa和0.58 GPa下记录的不同晶体数据合并后，得到了两组具有良好分辨率、完整性、冗余度和R(sym)值的结构振幅。在0.69 GPa下的第三组数据除完整性较低外，质量类似。这三种结构已得到优化。通过一系列衍射图像捕捉到了tHEWL晶体在超过0.82 GPa时压力诱导的晶体有序性丧失。
