the congenital cataract-linked g61c mutation destabilizes γd-crystallin and promotes non-native aggregation的先天性cataract-linked g61c突变造成γd-crystallin和促进非聚合.pdfVIP

The Congenital Cataract-Linked G61C Mutation Destabilizes cD-Crystallin and Promotes Non-Native Aggregation 1. 1. 2,3.¤ 1,4 2,3 1 Wang Zhang , Hong-Chen Cai , Fei-Feng Li , Yi-Bo Xi , Xu Ma *, Yong-Bin Yan * 1 State Key Laboratory of Biomembrane and Membrane Biotechnology, School of Life Sciences, Tsinghua University, Beijing, China, 2 Department of Genetics, National Research Institute for Family Planning, Beijing, China, 3 Peking Union Medical College, Tsinghua University, Beijing, China, 4 Institute of Biophysics, Lanzhou University, Lanzhou, China Abstract cD-crystallin is one of the major structural proteins in human eye lens. The solubility and stability of cD-crystallin play a crucial role in maintaining the optical properties of the lens during the life span of an individual. Previous study has shown that the inherited mutation G61C results in autosomal dominant congenital cataract. In this research, we studied the effects of the G61C mutation on cD-crystallin structure, stability and aggregation via biophysical methods. CD, intrinsic and extrinsic fluorescence spectroscopy indicated that the G61C mutation did not affect the native structure of cD-crystallin. The stability of cD-crystallin against heat- or GdnHCl-induced denaturation was significantly decreased by the mutation, while no influence was observed on the acid-induced unfolding. The mutation mainly affected the transition from the native state to the intermediate but not that from the intermediate to the unfolded or aggregated states. At high temperatures, both proteins were able to form aggregates, and the aggregation of the mutant was much more serious than the wild type protein at the same temperature. At body tempe