Heat-induced denaturation of cataract-related human γD-crystallin
Objective To reveal the thermally induced denaturation of wild-type human γD-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...
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Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College.
2025-01-01
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| Series: | Jichu yixue yu linchuang |
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| Online Access: | https://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2025-45-1-1.pdf |
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| author | ZHOU Xin, LI Zhenyan, LI Shuyuan, ZHANG Wenbo, WANG Chenxuan |
| author_facet | ZHOU Xin, LI Zhenyan, LI Shuyuan, ZHANG Wenbo, WANG Chenxuan |
| author_sort | ZHOU Xin, LI Zhenyan, LI Shuyuan, ZHANG Wenbo, WANG Chenxuan |
| collection | DOAJ |
| description | Objective To reveal the thermally induced denaturation of wild-type human γD-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 decreased 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. |
| format | Article |
| id | doaj-art-b31c392bd8b248dcb630430708fa7003 |
| institution | OA Journals |
| issn | 1001-6325 |
| language | zho |
| publishDate | 2025-01-01 |
| publisher | Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College. |
| record_format | Article |
| series | Jichu yixue yu linchuang |
| spelling | doaj-art-b31c392bd8b248dcb630430708fa70032025-08-20T01:50:33ZzhoInstitute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College.Jichu yixue yu linchuang1001-63252025-01-014511610.16352/j.issn.1001-6325.2025.01.0001Heat-induced denaturation of cataract-related human γD-crystallinZHOU Xin, LI Zhenyan, LI Shuyuan, ZHANG Wenbo, WANG Chenxuan0Department of Biophysics and Structural Biology, State Key Laboratory of Common Mechanism Research for Major Diseases, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC, Beijing 100005, ChinaObjective To reveal the thermally induced denaturation of wild-type human γD-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 decreased 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.https://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2025-45-1-1.pdfcrystallin|thermal stability|aggregation|intrinsic fluorescence|static light scattering |
| spellingShingle | ZHOU Xin, LI Zhenyan, LI Shuyuan, ZHANG Wenbo, WANG Chenxuan Heat-induced denaturation of cataract-related human γD-crystallin Jichu yixue yu linchuang crystallin|thermal stability|aggregation|intrinsic fluorescence|static light scattering |
| title | Heat-induced denaturation of cataract-related human γD-crystallin |
| title_full | Heat-induced denaturation of cataract-related human γD-crystallin |
| title_fullStr | Heat-induced denaturation of cataract-related human γD-crystallin |
| title_full_unstemmed | Heat-induced denaturation of cataract-related human γD-crystallin |
| title_short | Heat-induced denaturation of cataract-related human γD-crystallin |
| title_sort | heat induced denaturation of cataract related human γd crystallin |
| topic | crystallin|thermal stability|aggregation|intrinsic fluorescence|static light scattering |
| url | https://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2025-45-1-1.pdf |
| work_keys_str_mv | AT zhouxinlizhenyanlishuyuanzhangwenbowangchenxuan heatinduceddenaturationofcataractrelatedhumangdcrystallin |