Graduate School of Peking Union Medical College, Beijing, China.
Hum Mutat. 2012 Feb;33(2):391-401. doi: 10.1002/humu.21648. Epub 2011 Dec 8.
Congenital cataract is one of the leading causes of human blindness. In this study, we identified a novel, heterozygous c.385G<T mutation in CRYGC that resulted in the substitution of a highly conserved glycine by cysteine at codon 129 (p.Gly129Cys) in a three-generation Chinese family with autosomal dominant congenital nuclear cataract by sequencing candidate genes. Using zebrafish as a model, we demonstrated that γC-crystallin p.Gly129Cys mutant caused the vacuole and the incomplete denucleation of lens, recapitulating the cataract phenotype in human beings. Molecular modeling and spectroscopic studies indicated that the mutation impaired the tertiary structure of the protein by modifying the H-bonding network in the C-terminal domain. The mutation led to a dramatic decrease in the thermal stability of γC-crystallin, and a significant increase in the propensity of aggregation when subject to storage at high concentrations, heat, and UV- irradiation stresses. Taken together, these results indicate that a novel γC-crystallin p.Gly129Cys mutation impaired the tertiary structure of the protein and caused cataract formation, which provides a new insight into how the mutation may affect the γC-crystallin structure, stability, and function. Our study also highlighted zebrafish as a valuable model tool for studying congenital inherited cataract.
先天性白内障是导致人类失明的主要原因之一。在这项研究中,我们通过对一个三代常染色体显性遗传性先天性核白内障家系进行候选基因测序,发现了一个CRYGC 基因的新型杂合 c.385G>T 突变,导致 129 密码子的甘氨酸被半胱氨酸取代(p.Gly129Cys)。我们利用斑马鱼作为模型,证明 γC-晶状体蛋白 p.Gly129Cys 突变导致了空泡和晶状体不完全去核,重现了人类白内障的表型。分子建模和光谱研究表明,该突变通过修饰 C 末端结构域中的氢键网络,破坏了蛋白质的三级结构。该突变导致 γC-晶状体蛋白的热稳定性显著降低,在高浓度、热和 UV 照射等储存条件下,其聚集倾向显著增加。综上所述,这些结果表明,一种新型的 γC-晶状体蛋白 p.Gly129Cys 突变破坏了蛋白质的三级结构,导致白内障的形成,为突变如何影响 γC-晶状体蛋白的结构、稳定性和功能提供了新的见解。我们的研究还强调了斑马鱼作为研究先天性遗传性白内障的有价值的模型工具。