Peptide binding induces large scale changes in inter-domain mobility in human Pin1
Peptide binding induces large scale changes in inter-domain mobility in human Pin1
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DOI：
10.1074/jbc.m300796200
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发表时间：
2003-07-11
影响因子：
4.8
通讯作者：
Fiebig, KM
中科院分区：
文献类型：
--
作者：
Jacobs, DM;Saxena, K;Fiebig, KM
Pin1是细胞周期调控所必需的多肽-脯氨酰顺反异构酶(PPIase)。Pin1催化的肽基-Pro异构化提供了一个关键的构象开关来激活具有共同的磷酸化-丝氨酸/苏氨酸序列基序的磷酸化位点。这个基序在细胞对各种信号的反应中被普遍利用。Pin1能够在竞争相同底物的两个不同位置结合含有磷酸化丝氨酸/苏氨酸的序列。一个结合位点位于N-末端WW结构域，这是蛋白质靶向和定位所必需的。另一个结合部位位于PPIase的C端催化结构域，结构上与PPIase的FK506结合蛋白(FKBP)同源。12个残基的柔性连接子连接WW和催化结构域。为了表征全长Pin1在溶液中的结构和动力学，使用高分辨率核磁共振方法绘制了Pin1的两个结构域之间相互作用的性质。此外，还研究了目标多肽对结构域相互作用的影响。这些研究揭示了领域相互作用的动态图景。N-15自旋驰豫数据、差分化学位移映射和残余偶极耦合数据表明，Pin1既可以作为两个独立的结构域，通过柔性连接体连接，也可以作为一个完整的结构域，根据结合肽的序列进行一定程度的铰链弯曲运动。根据PIN1相互作用伙伴的数量，讨论了调节相对结构域灵活性的功能重要性。
Pin1 is a peptidyl-prolyl cis/trans isomerase (PPIase) essential for cell cycle regulation. Pin1-catalyzed peptidylprolyl isomerization provides a key conformational switch to activate phosphorylation sites with the common phospho-Ser/Thr-Pro sequence motif. This motif is ubiquitously exploited in cellular response to a variety of signals. Pin1 is able to bind phospho-Ser/Thr-Pro containing sequences at two different sites that compete for the same substrate. One binding site is located within the N-terminal WW domain, which is essential for protein targeting and localization. The other binding site is located in the C-terminal catalytic domain, which is structural homologous to the FK506-binding protein (FKBP) class of PPIases. A flexible linker of 12 residues connects the WW and catalytic domain. To characterize the structure and dynamics of full-length Pin1 in solution, high resolution NMR methods have been used to map the nature of interactions between the two domains of Pin1. In addition, the influence of target peptides on domain interactions has been investigated. The studies reveal a dynamic picture of the domain interactions. N-15 spin relaxation data, differential chemical shift mapping, and residual dipolar coupling data indicate that Pin1 can either behave as two independent domains connected by the flexible linker or as a single intact domain with some amount of hinge bending motion depending on the sequence of the bound peptide. The functional importance of the modulation of relative domain flexibility in light of the multitude of interaction partners of Pin1 is discussed.