Molecular basis of filamin A-FilGAP interaction and its impairment in congenital disorders associated with filamin A mutations.
Molecular basis of filamin A-FilGAP interaction and its impairment in congenital disorders associated with filamin A mutations.
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DOI：
10.1371/journal.pone.0004928
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发表时间：
2009
期刊：
影响因子：
3.7
通讯作者：
Stossel TP
中科院分区：
文献类型：
--
作者：
Nakamura F;Heikkinen O;Pentikäinen OT;Osborn TM;Kasza KE;Weitz DA;Kupiainen O;Permi P;Kilpeläinen I;Ylänne J;Hartwig JH;Stossel TP
细丝蛋白 A (FLNa) 是一种具有多个结合伙伴的重要细胞骨架蛋白，其突变会导致人类发育异常。我们确定了 FLNa 第 23 个 Ig 重复序列 (IgFLNa23) 与 FilGAP 相互作用的结构，FilGAP 是一种 Rac 特异性 GTP 酶激活蛋白，也是细胞极性和运动的调节因子，以及三种疾病相关突变对这种相互作用的影响。 NMR 结构分析和计算机建模相结合，揭示了 IgFLNa23 的 C 和 D β 链与 FilGAP C 端 32 个残基之间的结构界面细节。预测的关键界面残基的诱变证实了两种蛋白质之间的结合限制。生成特定功能丧失的 FLNa 构建体并用于分析体内 FLNa-FilGAP 相互作用的重要性。点突变揭示 FLNa-FilGAP 界面的破坏会扰乱细胞扩散。 FilGAP 不结合 FLNa 同源物 FLNb 或 FLNc，证实了这种相互作用对人类 FLNa 突变的重要性。紧密复合物的形成需要双方的二聚化和结合表面的正确对齐，这是由 FLNa 重复 23 和 24 之间的灵活铰链结构域促进的。与人类发育异常相关的 FLNa 突变会破坏结合相互作用，并削弱 FLNa/F-肌动蛋白网络在高机械应力下的弹性。根据结构进行突变分析可以生成用于探测 FLNa 特定细胞相互作用的试剂。疾病相关的 FLNa 突变对 FLNa 功能具有明显影响。
Mutations in filamin A (FLNa), an essential cytoskeletal protein with multiple binding partners, cause developmental anomalies in humans. We determined the structure of the 23rd Ig repeat of FLNa (IgFLNa23) that interacts with FilGAP, a Rac-specific GTPase-activating protein and regulator of cell polarity and movement, and the effect of the three disease-related mutations on this interaction. A combination of NMR structural analysis and in silico modeling revealed the structural interface details between the C and D β-strands of the IgFLNa23 and the C-terminal 32 residues of FilGAP. Mutagenesis of the predicted key interface residues confirmed the binding constraints between the two proteins. Specific loss-of-function FLNa constructs were generated and used to analyze the importance of the FLNa-FilGAP interaction in vivo. Point mutagenesis revealed that disruption of the FLNa-FilGAP interface perturbs cell spreading. FilGAP does not bind FLNa homologs FLNb or FLNc establishing the importance of this interaction to the human FLNa mutations. Tight complex formation requires dimerization of both partners and the correct alignment of the binding surfaces, which is promoted by a flexible hinge domain between repeats 23 and 24 of FLNa. FLNa mutations associated with human developmental anomalies disrupt the binding interaction and weaken the elasticity of FLNa/F-actin network under high mechanical stress. Mutational analysis informed by structure can generate reagents for probing specific cellular interactions of FLNa. Disease-related FLNa mutations have demonstrable effects on FLNa function.
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影响因子：
30.8
作者：
Krakow, D;Robertson, SP;Cohn, DH
通讯作者：
Cohn, DH
影响因子：
16.2
作者：
Fox, JW;Lamperti, ED;Walsh, CA
通讯作者：
Walsh, CA
影响因子：
4.8
作者：
Nakamura, F;Hartwig, JH;Szymanski, PT
通讯作者：
Szymanski, PT
影响因子：
5.6
作者：
Herrmann, T;Güntert, P;Wüthrich, K
通讯作者：
Wüthrich, K
DOI：
10.1083/jcb.200707073
发表时间：
2007-12-03
期刊：
The Journal of cell biology
影响因子：
--
作者：
Nakamura F;Osborn TM;Hartemink CA;Hartwig JH;Stossel TP
通讯作者：
Stossel TP