Structure of cysteine- and glycine-rich protein CRP2 -: Backbone dynamics reveal motional freedom and independent spatial orientation of the LIM domains
Structure of cysteine- and glycine-rich protein CRP2 -: Backbone dynamics reveal motional freedom and independent spatial orientation of the LIM domains
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DOI：
10.1074/jbc.273.36.23233
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发表时间：
1998-09-04
影响因子：
4.8
通讯作者：
Bister, K
中科院分区：
文献类型：
--
作者：
Konrat, R;Kräutler, B;Bister, K
富含半胱氨酸和甘氨酸的蛋白质家族（CRP 1、CRP 2和CRP 3）的成员含有两个锌结合LIM结构域，LIM1（氨基末端）和LIM2（羧基末端），并且涉及与分化、生长控制和发病机制相关的多种细胞过程。在这里，我们报告的解决方案的全长重组鹌鹑CRP 2的结构确定多维三重共振NMR光谱。结构分析表明，在全长蛋白的上下文中的两个LIM结构域的全局折叠与最近确定的鹌鹑CRP 2的分离的单个LIM结构域的溶液结构相同，两个结构域的相对空间取向没有偏好。这支持了这两个LIM结构域是CRP蛋白的独立结构和推测功能模块的观点。这也反映在由（15）N弛豫值（T（1）、T（2）和核Overhauser效应）探测的CRP 2的动力学性质上。无模型分析揭示了本地变化的流动性沿着骨干的两个LIM结构域在天然蛋白质，类似于那些观察到的孤立的结构域。有趣的是，在两个LIM结构域之间的58个氨基酸的接头区域中观察到的快速和缓慢运动赋予CRP 2广泛的运动自由度。动态分析表明两个LIM结构域的独立骨架移动性，并排除了全长CRP 2中相关的LIM结构域运动。蛋白质中包含多个LIM基序的LIM结构域在结构上和动力学上彼此独立的发现支持了这些蛋白质可以作为衔接分子，将两种或多种蛋白质组分排列成大分子复合物。
Members of the cysteine- and glycine-rich protein family (CRP1, CRP2, and CRP3) contain two zinc-binding LIM domains, LIM1 (amino-terminal) and LIM2 (carboxyl-terminal), and are implicated in diverse cellular processes linked to differentiation, growth control, and pathogenesis. Here we report the solution structure of full-length recombinant quail CRP2 as determined by multi-dimensional triple-resonance NMR spectroscopy. The structural analysis revealed that the global fold of the two LIM domains in the context of the full-length protein is identical to the recently determined solution structures of the isolated individual LIM domains of quail CRP2, There is no preference in relative spatial orientation of the two domains. This supports the view that the two LIM domains are independent structural and presumably functional modules of CRP proteins. This is also reflected by the dynamic properties of CRP2 probed by (15)N relaxation values (T(1), T(2), and nuclear Overhauser effect). A model-free analysis revealed local variations in mobility along the backbone of the two LIM domains in the native protein, similar to those observed for the isolated domains. Interestingly, fast and slow motions observed in the 58-amino acid linker region between the two LIM domains endow extensive motional freedom to CRP2, The dynamic analysis indicates independent backbone mobility of the two LIM domains and rules out correlated LIM domain motion in full-length CRP2, The finding that the LIM domains in a protein encompassing multiple LIM motifs are structurally and dynamically independent from each other supports the notion that these proteins may function as adaptor molecules arranging two or more protein constituents into a macromolecular complex.