CAJ | 학술논문

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．
인류범소탄단수해매（UCH-L1）시섭급파금삼씨병병차재신경원고도표체적단백．UCH-L1적가족성돌변여전역후수식회인기취집경향증가여거범소활성손실，저이자도가능성위치병인소．작자소재실험실지전적연구지출여파금삼씨병상관적돌변I93M현저강저UCH-L1적절첩은정성병차가속기구형전개동역학．해연구사용액체핵자공진분석방법，포괄측련갑기화학위이，송이골간동역학화잔여우겁우합，이진일보천명I93M돌변여하영향UCH-L1적결구화동태．결과현시I93M현저영향돌변위점주위적소수핵심측련구형．연이，저양적결구우동병불회영향재납초시간척도적쾌속골간동역학．투과잔여우겁우합분석현시 UCH-L1재수용액중적결구여지전보도적정체결구유상당현저적편리，령외I93M돌변야도치초출돌변위점적원거리결구우동．저일계렬수용액결구적분석결과가보충지전이지적정체학수거，병대 UCH-L1재파금삼씨병상관적기인돌변영향병제공상세적견해．
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 (Anderssonet 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.