Heat-induced denaturation of cataract-related human γ D-crystallin
10.16352/j.issn.1001-6325.2025.01.0001
- VernacularTitle:白内障相关人γD晶状体蛋白的热诱导变性
- Author:
Xin ZHOU
1
;
Zhenyan LI
;
Shuyuan LI
;
Wenbo ZHANG
;
Chenxuan WANG
Author Information
1. 中国医学科学院基础医学研究所 北京协和医学院基础学院 生物物理及结构生物学系重大疾病共性机制研究全国重点实验室,北京 100005
- Keywords:
crystallin;
thermal stability;
aggregation;
intrinsic fluorescence;
static light scattering
- From:
Basic & Clinical Medicine
2025;45(1):1-6
- CountryChina
- Language:Chinese
-
Abstract:
Objective To reveal the thermally induced denaturation of wild-type human γ D-crystallin(HGD)and congenital cataract-related mutant(HGD P23T),and compare the differences in the structural changes between wild-type and mutants during a heating process.Methods HGD and HGD P23T were expressed and purified.The temperature-dependent intrinsic fluorescence intensity and static light scattering intensity of the protein samples were measured to reveal the temperature-dependent folding and aggregation structural changes of HGD and HGD P23T.Results When the temperature was below 70℃,the barycentric mean of the intrinsic fluorescence of HGD and HGD P23T shifted towards a longer wavelength with increasing temperature and the fluorescence intensity de-creased indicating the unfolded protein conformations.The conformational stability of HGD P23T was weaker than that of HGD.When temperature was higher than 70℃,the static light scattering intensity increased significantly with temperature,indicating protein aggregation upon heating.Relative to the wild-type,HGD P23T showed a stronger aggregation potency.Conclusions Heating disrupts the folding conformation of Γd-crystallin,induces the unfolded protein to aggregate.The disease-associated P23T mutation significantly reduces the conformational stability of Γd-crystallin.
