Structure of the S100A6 complex with a fragment from the C-terminal domain of Siah-1 interacting protein: a novel mode for S100 protein target recognition.
Structure of the S100A6 complex with a fragment from the C-terminal domain of Siah-1 interacting protein: a novel mode for S100 protein target recognition.
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DOI：
10.1021/bi801233z
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发表时间：
2008-10-14
期刊：
影响因子：
2.9
通讯作者：
Chazin, Walter J.
中科院分区：
文献类型：
--
作者：
Lee, Young-Tae;Dimitrova, Yoana N.;Schneider, Gabriela;Ridenour, Whitney B.;Bhattacharya, Shibani;Soss, Sarah E.;Caprioli, Richard M.;Filipek, Anna;Chazin, Walter J.
S100A6 是 Ca2+ 结合 EF-hand 蛋白 S100 亚家族的成员，已被证明与钙周期蛋白结合蛋白/Siah-1 相互作用蛋白 (CacyBP/SIP；SIP) 相互作用，后者是在基因毒性应激下形成的 SCF 样 E3 连接酶复合物 (SCF-TBL1) 的亚基。 SIP 在该复合体中充当支架，将 E2 招募模块 Siah-1 与底物招募模块 Skp1-TBL1 连接起来。基于细胞的功能测定表明 S100A6 调节 SCF-TBL1 的活性。如果能够选择性抑制 S100A6-SIP 相互作用而不干扰这两种蛋白质的任何其他功能，则可以增强基于细胞的实验结果。为此，通过 NMR 在溶液中测定了 S100A6-SIP 复合物的结构，并通过等温滴定量热法表征了相互作用的强度。在初始步骤中，SIP 中 S100A6 的最小结合区域被映射到 C 端结构域中的 31 个残基片段 (Ser189-Arg219)。 S100A6-SIP(189-219) 复合物的结构表明，SIP(189-219) 形成两个螺旋，其中第一个螺旋 (Met193-Tyr200) 以规范结合模式与 S100A6 相互作用。第二个螺旋 (Met207-Val216) 位于 S100A6 二聚体界面上方，这是一种与 S100A6 结合的模式，以前从未在与 S100 蛋白结合的任何靶标中观察到这种模式。一系列基于结构的 SIP 突变显示 S100A6 结合亲和力降低，为 S100A6-SIP 相互作用的直接功能分析奠定了基础。
S100A6 is a member of the S100 subfamily of Ca2+ binding EF-hand proteins that has been shown to interact with calcyclin binding protein/Siah-1 interacting protein (CacyBP/SIP; SIP), a subunit of an SCF-like E3 ligase complex (SCF-TBL1) formed under genotoxic stress. SIP serves as a scaffold in this complex, linking the E2-recruiting module Siah-1 to the substrate-recruiting module Skp1-TBL1. A cell-based functional assay suggests that S100A6 modulates the activity of SCF-TBL1. The results from the cell-based experiments could be enhanced if it were possible to selectively inhibit S100A6-SIP interactions without perturbing any other functions of the two proteins. To this end, the structure of the S100A6-SIP complex was determined in solution by NMR and the strength of the interaction was characterized by isothermal titration calorimetry. In an initial step, the minimal binding region in SIP for S100A6 was mapped to a 31 residue fragment (Ser189-Arg219) in the C-terminal domain. The structure of the S100A6-SIP(189–219) complex revealed that SIP(189–219) forms two helices, the first of which (Met193-Tyr200) interacts with S100A6 in a canonical binding mode. The second helix (Met207-Val216) lies over the S100A6 dimer interface, a mode of binding to S100A6 that has not previously been observed for any target bound to an S100 protein. A series of structure-based SIP mutations showed reduced S100A6 binding affinity, setting the stage for direct functional analysis of S100A6-SIP interactions.
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影响因子：
8
作者：
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通讯作者：
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影响因子：
2.9
作者：
Bhattacharya, S;Lee, YT;Chazin, WJ
通讯作者：
Chazin, WJ
DOI：
10.1073/pnas.81.19.6004
发表时间：
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期刊：
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