Structure of PICK1 and other PDZ domains obtained with the help of self-binding C-terminal extensions
Structure of PICK1 and other PDZ domains obtained with the help of self-binding C-terminal extensions
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DOI：
10.1110/ps.062657507
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发表时间：
2007-04-01
期刊：
影响因子：
8
通讯作者：
Doyle, Declan A.
中科院分区：
文献类型：
--
作者：
Elkins, Jonathan M.;Papagrigoriou, Evangelos;Doyle, Declan A.
PDZ结构域是蛋白质-蛋白质相互作用模块，其通常与其靶蛋白的C末端结合。PDZ结构域的预期结合配偶体的C末端四个氨基酸通常是结合特异性的决定因素。为了确定许多PDZ结构域的结构，我们在PDZ结构域截短突变体的C末端包括了适当的四个残基延伸，设计用于自结合。产生每个PDZ结构域的多个截短。四个残基延伸，这代表了已知的特异性序列的目标PDZ结构域，并涵盖了I类和II类基序，形成分子间接触的预期方式的PDZ结构域与蛋白C末端的相互作用，为两个类。我们展示了使用这种方法结晶的八个独特PDZ结构域的结构，并重点关注四个提供选择性（PICK 1和DLG 2的第三个PDZ结构域）、结合位点灵活性（MPDZ的第三个PDZ结构域）和肽结构域相互作用（MPDZ第12个PDZ结构域）的信息。对我们的结果的分析显示，与没有C-末端四个残基延伸的PDZ结构域的类似截短相比，通过使用这种方法获得PDZ结构域晶体的机会明显提高。
PDZ domains are protein-protein interaction modules that generally bind to the C termini of their target proteins. The C-terminal four amino acids of a prospective binding partner of a PDZ domain are typically the determinants of binding specificity. In an effort to determine the structures of a number of PDZ domains we have included appropriate four residue extensions on the C termini of PDZ domain truncation mutants, designed for self-binding. Multiple truncations of each PDZ domain were generated. The four residue extensions, which represent known specificity sequences of the target PDZ domains and cover both class I and II motifs, form intermolecular contacts in the expected manner for the interactions of PDZ domains with protein C termini for both classes. We present the structures of eight unique PDZ domains crystallized using this approach and focus on four which provide information on selectivity (PICK1 and the third PDZ domain of DLG2), binding site flexibility (the third PDZ domain of MPDZ), and peptide-domain interactions (MPDZ 12th PDZ domain). Analysis of our results shows a clear improvement in the chances of obtaining PDZ domain crystals by using this approach compared to similar truncations of the PDZ domains without the C- terminal four residue extensions.