Structural Perturbation of the Parkinson´s Disease-Associated I93M Mutation in Human UCH-L1 Revealed by Solution State NMR Spectroscopy

doi:10.11938/cjmr20150215

Abstract

Abstract:

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.

Key words: Parkinson´s disease, UCH-L1, protein folding, solution state NMR spectroscopy, chemical shift perturbation, order parameter, residual dipolar coupling

CLC Number:

O482.53

KUMAR Sriramoju M , LYU Ping-chiang, HSU Shang-te Danny, . Structural Perturbation of the Parkinson´s Disease-Associated I93M Mutation in Human UCH-L1 Revealed by Solution State NMR Spectroscopy[J]. Chinese Journal of Magnetic Resonance, 2015, 32(2): 329-341.

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