The C-terminal V5 domain of Protein Kinase Cα is intrinsically disordered, with propensity to associate with a membrane mimetic.
The C-terminal V5 domain of Protein Kinase Cα is intrinsically disordered, with propensity to associate with a membrane mimetic.
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DOI：
10.1371/journal.pone.0065699
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Igumenova TI
中科院分区：
文献类型：
--
作者：
Yang Y;Igumenova TI
C端V5结构域是蛋白激酶C异构体（PKC）中最可变的结构域之一。V5通过与同种型特异性衔接蛋白的相互作用以及可能通过与其他PKC结构域的特异性分子内相互作用赋予其亲本酶同种型特异性。溶液中V5区域的结构信息是稀疏的。本工作的目的是确定PKC α亚型（V5α）的V5结构域的构象偏好性，并评估其与膜模拟物结合的能力。我们发现V5α及其磷酸化模拟变体dmV 5 α本质上是无序的蛋白质结构域。磷酸化模拟突变不改变多肽骨架的整体构象，如化学位移扰动的局部性质和二级结构倾向评分所证明的。然而，Thr 638-Pro639-Pro640转角基序的“顺-反”构象异构体的数量在dmV 5 α中增加，沿着转角和疏水基序之间区域的构象灵活性，这与通过肽基脯氨酰异构酶Pin 1下调PKCα有关。野生型和dmV 5 α均与由两性离子去污剂n-十二烷基磷酸胆碱制成的胶束缔合。在胶束结合时，V5α获得在保守的“NFD”基序和整个C-末端三分之一结构域处形成螺旋结构的更高倾向。V5α进入疏水胶束环境的能力表明它可能在PKC成熟过程中作为膜锚。
The C-terminal V5 domain is one of the most variable domains in Protein Kinase C isoforms (PKCs). V5 confers isoform specificity on its parent enzyme through interactions with isoform-specific adaptor proteins and possibly through specific intra-molecular interactions with other PKC domains. The structural information about V5 domains in solution is sparse. The objective of this work was to determine the conformational preferences of the V5 domain from the α isoform of PKC (V5α) and evaluate its ability to associate with membrane mimetics. We show that V5α and its phosphorylation-mimicking variant, dmV5α, are intrinsically disordered protein domains. Phosphorylation-mimicking mutations do not alter the overall conformation of the polypeptide backbone, as evidenced by the local nature of chemical shift perturbations and the secondary structure propensity scores. However, the population of the “cis-trans” conformer of the Thr638-Pro639-Pro640 turn motif, which has been implicated in the down-regulation of PKCα via peptidyl-prolyl isomerase Pin1, increases in dmV5α, along with the conformational flexibility of the region between the turn and hydrophobic motifs. Both wild type and dmV5α associate with micelles made of a zwitterionic detergent, n-dodecylphosphocholine. Upon micelle binding, V5α acquires a higher propensity to form helical structures at the conserved “NFD” motif and the entire C-terminal third of the domain. The ability of V5α to partition into the hydrophobic micellar environment suggests that it may serve as a membrane anchor during the PKC maturation process.
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