The dominant deafness point mutation GJB2 p.D179N affects the oligomeric equilibrium of connexin 26
10.3969/j.issn.1006-7299.2025.03.011
- VernacularTitle:显性致聋突变GJB2 p.D179N影响缝隙连接蛋白26寡聚平衡状态
- Author:
Huahong LUO
1
;
Guohui HUANG
;
Yunge GAO
;
Tao YANG
;
Hao WU
;
Zhihua ZHANG
Author Information
1. 上海交通大学医学院附属第九人民医院耳鼻咽喉头颈外科(上海 200011);上海市耳鼻疾病转化医学重点实验室(上海 200125);上海交通大学医学院耳科学研究所(上海 200125)
- Publication Type:Journal Article
- Keywords:
Deafness mutation;
Connexin 26;
Protein purification;
Size-exclusion chromatography;
Cryo-electron microscopy single-particle analysis
- From:
Journal of Audiology and Speech Pathology
2025;33(3):259-264
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the impact of the dominant deafness point mutation p.D179N on the oli-gomeric equilibrium state of Connexin 26(Cx26).Methods The wild-type Cx26 fusion protein(Cx26-WT-GFP)and mutant fusion proteins(Cx26-D179N-GFP,Cx26-D179C-GFP)were expressed in HEK293F cells.By using Fluorescence-detection size-exclusion chromatography(FSEC)and size-exclusion chromatography(SEC)to analysis the oligomeric state of the target protein based on malecular weight under the condition of solubilization and purifica-tion respectively.Cryo-electron microscopy(Cryo-EM)single particle analysis(SPA)was conducted to analysis the target protein's oligomeric states based on the 2D classification morphology of the protein particles.Results In vitro,the wild-type Cx26 protein(Cx26-WT)is almost exclusively dodecameric.The deafness mutation p.D179N protein(Cx26-D179N)exists as both dodecamers and hexamers,whereas the artificial mutation p.D179C protein(Cx26-D179C)does not form dodecamers.Conclusion The dominant deafness mutation GJB2 p.D179N could weaken the ability of docking between hexameric proteins,which could affect the balance between hexamers and do-decamers.
