Two-dimensional protein crystallization via metal-ion coordination by naturally occurring surface histidines.
Two-dimensional protein crystallization via metal-ion coordination by naturally occurring surface histidines.
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DOI：
10.1073/pnas.93.10.4937
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发表时间：
1996-05
影响因子：
11.1
通讯作者：
W. Frey;W. Schief;D. W. Pack;C. T. Chen;A. Chilkoti;P. Stayton;V. Vogel;F. H. Arnold
中科院分区：
文献类型：
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作者：
W. Frey;W. Schief;D. W. Pack;C. T. Chen;A. Chilkoti;P. Stayton;V. Vogel;F. H. Arnold
提出了一种通过表面组氨酸残基与脂质螯合的二价金属离子配位来靶向和结晶脂质界面上的蛋白质的有效的和潜在的通用方法。这种方法应该适用于各种天然或工程蛋白质的结晶，这里以链霉亲和素在亚氨基二乙酸酯-铜(II)脂质单层上的结晶为例说明了这一点。这种方法允许控制蛋白质在界面上的取向，这对于方便地生产高度有序的蛋白质阵列和在二维晶体结构分析中绘制电子密度图具有重要意义。天然链霉亲和素通过位于生物素结合口袋附近的蛋白质表面的His-87与亚氨基二乙酸酯-铜脂结合。二维链霉亲和素晶体显示出一种以前未描述的微观形状，不同于在生物素化脂质下形成的晶体。
A powerful and potentially general approach to the targeting and crystallization of proteins on lipid interfaces through coordination of surface histidine residues to lipid-chelated divalent metal ions is presented. This approach, which should be applicable to the crystallization of a wide range of naturally occurring or engineered proteins, is illustrated here by the crystallization of streptavidin on a monolayer of an iminodiacetate-Cu(II) lipid spread at the air-water interface. This method allows control of the protein orientation at interfaces, which is significant for the facile production of highly ordered protein arrays and for electron density mapping in structural analysis of two-dimensional crystals. Binding of native streptavidin to the iminodiacetate-Cu lipids occurs via His-87, located on the protein surface near the biotin binding pocket. The two-dimensional streptavidin crystals show a previously undescribed microscopic shape that differs from that of crystals formed beneath biotinylated lipids.