Department of Ophthalmology, The Second Affiliated Hospital, Zhejiang University School of Medicine, 88 Jiefang Road, Hangzhou 310030, China.
Department of Ophthalmology, Zhejiang Provincial People's Hospital, Affiliated People's Hospital, Hangzhou Medical College, 158 Shangtang Road, Hangzhou 310053, China.
Biomolecules. 2023 May 19;13(5):864. doi: 10.3390/biom13050864.
Congenital cataracts account for approximately 5-20% of childhood blindness worldwide and 22-30% of childhood blindness in developing countries. Genetic disorders are the primary cause of congenital cataracts. In this work, we investigated the underlying molecular mechanism of G149V point missense mutation in βB2-crystallin, which was first identified in a three-generation Chinese family with two affected members diagnosed with congenital cataracts. Spectroscopic experiments were performed to determine the structural differences between the wild type (WT) and the G149V mutant of βB2-crystallin. The results showed that the G149V mutation significantly changed the secondary and tertiary structure of βB2-crystallin. The polarity of the tryptophan microenvironment and the hydrophobicity of the mutant protein increased. The G149V mutation made the protein structure loose and the interaction between oligomers was reduced, which decreased the stability of the protein. Furthermore, we compared βB2-crystallin WT and the G149V mutant with their biophysical properties under environmental stress. We found that the G149V mutation makes βB2-crystallin more sensitive to environmental stresses (oxidative stress, UV irradiation, and heat shock) and more likely to aggregate and form precipitation. These features might be important to the pathogenesis of βB2-crystallin G149V mutant related to congenital cataracts.
先天性白内障约占全球儿童失明的 5-20%,发展中国家儿童失明的 22-30%。遗传疾病是先天性白内障的主要原因。在这项工作中,我们研究了首次在一个三代中国家庭中发现的βB2-晶体蛋白 G149V 点错义突变的潜在分子机制,该家庭中有两名受影响成员被诊断为先天性白内障。进行了光谱实验以确定野生型(WT)和βB2-晶体蛋白 G149V 突变体之间的结构差异。结果表明,G149V 突变显著改变了βB2-晶体蛋白的二级和三级结构。色氨酸微环境的极性和突变蛋白的疏水性增加。G149V 突变使蛋白质结构松散,寡聚物之间的相互作用减少,从而降低了蛋白质的稳定性。此外,我们比较了βB2-晶体蛋白 WT 和 G149V 突变体及其在环境胁迫下的生物物理特性。我们发现,G149V 突变使βB2-晶体蛋白对环境应激(氧化应激、紫外线照射和热休克)更敏感,更容易聚集并形成沉淀。这些特征对于与先天性白内障相关的βB2-晶体蛋白 G149V 突变体的发病机制可能很重要。