Crystallography and protein-protein interactions: biological interfaces and crystal contacts
Crystallography and protein-protein interactions: biological interfaces and crystal contacts
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DOI：
10.1042/bst0361438
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发表时间：
2008-12-01
影响因子：
3.9
通讯作者：
Forwood, Jade K.
中科院分区：
文献类型：
--
作者：
Kobe, Bostjan;Guncar, Gregor;Forwood, Jade K.
结晶学是研究蛋白质-蛋白质复合体结构的常用方法。然而，晶体结构并不能定义一个独特的蛋白质-蛋白质界面，区分“生物界面”和“晶体接触”通常也不是一帆风顺的。有一些计算方法可以区分它们，但它们的错误率很高，强调需要使用互补的结构和功能方法获得更多关于生物界面的数据。除了回顾解决这个问题的计算和实验方法外，我们还强调了两个相关的例子。我们实验室的第一个例子涉及酰基辅酶A硫代酯酶7的结构，其中这个双域蛋白的每个结构域都被单独结晶，但都产生了一个非功能性组装。通过化学交联法、分析性超速离心法和诱变法等多种互补方法，发现了全长蛋白质的结构。第二个例子涉及血小板膜糖蛋白Ibα-凝血酶复合体。两个小组报道了该络合物的晶体结构，但两种结构之间的所有相互作用界面都不同。我们的计算分析没有完全解决差异的原因，但提供了对系统的有趣见解。这篇综述强调了用互补的实验和计算方法来补充结晶学研究的必要性。
Crystallography is commonly used for studying the structures of protein-protein complexes. However, a crystal structure does not define a unique protein-protein interface, and distinguishing a 'biological interface' from 'crystal contacts' is often not straightforward. A number of computational approaches exist for distinguishing them, but their error rate is high, emphasizing the need to obtain further data on the biological interface using complementary structural and functional approaches. in addition to reviewing the computational and experimental approaches for addressing this problem, we highlight two relevant examples. The first example from our laboratory involves the structure of acyl-CoA thioesterase 7, where each domain of this two-domain protein was crystallized separately, but both yielded a non-functional assembly. The structure of the full-length protein was uncovered using a combination of complementary approaches including chemical cross-linking, analytical ultracentrifugation and mutagenesis. The second example involves the platelet glycoprotein Ib alpha-thrombin complex. Two groups reported the crystal structures of this complex, but all the interacting interfaces differed between the two structures. Our computational analysis did not fully resolve the reasons for the discrepancies, but provided interesting insights into the system. This review highlights the need to complement crystallographic studies with complementary experimental and computational approaches.