Structural characterization of the N-terminal kinase-interacting domain of an Hsp90-cochaperone Cdc37 by CD and solution NMR spectroscopy.
Structural characterization of the N-terminal kinase-interacting domain of an Hsp90-cochaperone Cdc37 by CD and solution NMR spectroscopy.
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DOI：
10.1016/j.bbapap.2019.06.007
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发表时间：
2019-09
期刊：
影响因子：
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通讯作者：
F. Ihama;Mami Yamamoto;C. Kojima;T. Fujiwara;K. Matsuzaki;Y. Miyata;M. Hoshino
中科院分区：
文献类型：
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作者：
F. Ihama;Mami Yamamoto;C. Kojima;T. Fujiwara;K. Matsuzaki;Y. Miyata;M. Hoshino
Cdc 37是一种靶向蛋白激酶的分子伴侣，它与Hsp 90协同作用，协助多种信号激酶的折叠、组装和成熟。它由三个不同的结构域组成：N-末端、中间和C-末端结构域。虽然中间结构域是Hsp 90结合结构域，但N-末端结构域被认为是激酶相互作用结构域。N-末端结构域在位置13处含有高度保守的Ser残基，并且该位点处的磷酸化已被证明是Cdc 37与信号激酶之间相互作用的先决条件。虽然Ser 13的磷酸化可能引起Cdc 37分子的一些构象变化，但对Cdc 37的N端结构域的结构知之甚少。我们研究了几个片段的蛋白质对应的N-末端区域的Cdc 37的圆二色谱和溶液NMR光谱的结构和动力学特性。我们发现Cdc 37的N端结构域呈现高度的动态结构，它处于α螺旋和更无序结构之间的平衡状态.我们还发现，Ser 13的磷酸化并没有显着改变Cdc 37的N-末端片段蛋白的整体结构。结果表明，可能需要更复杂的机制来解释Cdc 37与各种激酶的磷酸化激活相互作用。
Cdc37 is a protein kinase-targeting molecular chaperone, which cooperates with Hsp90 to assist the folding, assembly and maturation of various signaling kinases. It consists of three distinct domains: the N-terminal, middle, and C-terminal domain. While the middle domain is an Hsp90-binding domain, the N-terminal domain is recognized as a kinase-interacting domain. The N-terminal domain contains a well-conserved Ser residue at position 13, and the phosphorylation at this site has been shown to be a prerequisite for the interaction between Cdc37 and signaling kinases. Although the phosphorylation of Ser13 might induce some conformational change in Cdc37 molecule, little is known about the structure of the N-terminal domain of Cdc37. We examined the structural and dynamic properties of several fragment proteins corresponding to the N-terminal region of Cdc37 by circular dichroism and solution NMR spectroscopy. We found that the N-terminal domain of Cdc37 exhibits highly dynamic structure, and it exists in the equilibrium between α-helical and more disordered structures. We also found that phosphorylation at Ser13 did not significantly change the overall structure of N-terminal fragment protein of Cdc37. The results suggested that more complicated mechanisms might be necessary to explain the phosphorylation-activated interaction of Cdc37 with various kinases.