以液体核磁共振波谱分析与帕金森氏病相关的I93M 突变对人类泛素碳端水解酶结构的影响
1. 台湾清华大学 生物信息学和结构生物学研究所，台湾 新竹 30013；2. 生物化学研究所，台湾 台北 11529；3. 台湾大学 生物化学研究所，台湾 台北 10617
收稿日期: 2015-03-02
修回日期: 2015-05-08
网络出版日期: 2015-06-05
基金资助
Structural Perturbation of the Parkinson´s Disease-Associated I93M Mutation in Human UCH-L1 Revealed by Solution State NMR Spectroscopy
1. Institute of Bioinformatics and Structural Biology, Tsing Hua University of Taiwan, Hsinchu 30013, China; 2. Institute of Biological Chemistry, Taipei 11529, China; 3. Institute of Biochemical Science, Taiwan University, Taipei 10617, China
Received date: 2015-03-02
Revised date: 2015-05-08
Online published: 2015-06-05
Supported by
人类泛素碳端水解酶（UCH-L1）是涉及帕金森氏病并且在神经元高度表达的蛋白．UCH-L1 的家族性突变与转译后修饰会引起聚集倾向增加与去泛素活性损失，这二者都可能成为致病因素．作者所在实验室之前的研究指出与帕金森氏病相关的突变I93M 显著降低UCH-L1 的折叠稳定性并且加速其构型展开动力学．该研究使用液体核磁共振分析方法，包括侧链甲基化学位移，松弛骨干动力学和残余偶极耦合，以进一步阐明I93M 突变如何影响UCH-L1 的结构和动态．结果显示I93M 显著影响突变位点周围的疏水核心侧链构型．然而，这样的结构扰动并不会影响在纳秒时间尺度的快速骨干动力学．透过残余偶极耦合分析显示UCH-L1 在水溶液中的结构与之前报道的晶体结构有相当显著的偏离，另外I93M 突变也导致超出突变位点的远距离结构扰动．这一系列水溶液结构的分析结果可补充之前已知的晶体学数据，并对UCH-L1 在帕金森氏病相关的基因突变影响并提供详细的见解．
KUMAR Sriramoju M 1 , 呂平江1 , 徐尚德1 , 2 , 3* . 以液体核磁共振波谱分析与帕金森氏病相关的I93M 突变对人类泛素碳端水解酶结构的影响[J]. 波谱学杂志, 2015 , 32(2) : 329 -341 . DOI: 10.11938/cjmr20150215
Human ubiquitin C-terminal hydrolase, UCH-L1, is a highly abundant neuronal protein that is implicated in Parkinson´s disease (PD). Familial mutations and post-translational modifications of UCH-L1 have been reported to cause increased aggregation propensity and loss of de-ubiquitination activity, both of which may be pathogenic. We have recently demonstrated that a PD-associated mutation of UCH-L1, namely I93M, significantly destabilizes the folding
stability and accelerates the unfolding kinetics (Andersson et al. J Mol Biol, 2011, 407: 261－272). Here we report the use of solution state NMR spectroscopy, including side-chain methyl chemical shift, backbone relaxation dynamics and residual dipolar coupling (RDC) analyses, to further elucidate how the I93M mutation affects the structure and dynamics of UCH-L1. The results revealed altered side-chain packing within the hydrophobic core around the mutation site. However, such structural perturbation does not affect the fast backbone dynamics on the ns timescale. Furthermore, comparative RDC analysis suggests that the solution structure of UCH-L1 deviates considerably from the reported crystal structure and that the I93M mutation results in long-range structural perturbations far beyond the mutation site. These solution state-based structural findings complement previously reported crystallographic data to provide detailed insights into the impacts of the PD-associated mutation on UCH-L1.
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