title: The congenital cataract-linked G61C mutation destabilizes γD-crystallin and promotes non-native aggregation.
abstract: γD-crystallin is one of the major structural proteins in human eye lens. The solubility and stability of γD-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 γD-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 γD-crystallin. The stability of γD-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 temperature and acidic conditions, the mutant was more prone to form amyloid-like fibrils. The aggregation-prone property of the mutant was not altered by the addition of reductive reagent. These results suggested that the decrease in protein stability followed by aggregation-prone property might be the major cause in the hereditary cataract induced by the G61C mutation.
authors.affiliation: State Key Laboratory of Biomembrane and Membrane Biotechnology, School of Life Sciences, Tsinghua University, Beijing, China.
authors.forename: Wang
authors.initials: W
authors.lastname: Zhang
authors.forename: Hong-Chen
authors.initials: HC
authors.lastname: Cai
authors.forename: Fei-Feng
authors.initials: FF
authors.lastname: Li
authors.forename: Yi-Bo
authors.initials: YB
authors.lastname: Xi
authors.forename: Xu
authors.initials: X
authors.lastname: Ma
authors.forename: Yong-Bin
authors.initials: YB
authors.lastname: Yan
mesh_terms: D002386:Cataract / Q000235:genetics*; D003459:Crystallins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D011489:Protein Denaturation
chemical_list: D003459:Crystallins