Biochemical characterization of G64W mutant of acidic beta-crystallin 4

Wenqian Li; Qingshan Ji; Zhongjie Wei; Yu-Lei Chen; Zhiyong Zhang; Xueying Yin; Samaneh Khodi Aghmiuni; Muziying Liu; Weirong Chen; Lei Shi; Quan Chen; Xinzheng Du; Li Yu; Min-Jie Cao; Zhulou Wang; Shaohui Huang; Tengchuan Jin; Qiwei Wang

doi:10.1016/j.exer.2019.107712

Biochemical characterization of G64W mutant of acidic beta-crystallin 4

Exp Eye Res. 2019 Sep:186:107712. doi: 10.1016/j.exer.2019.107712. Epub 2019 Jun 26.

Authors

Wenqian Li¹, Qingshan Ji², Zhongjie Wei³, Yu-Lei Chen⁴, Zhiyong Zhang⁵, Xueying Yin⁵, Samaneh Khodi Aghmiuni⁵, Muziying Liu⁵, Weirong Chen⁶, Lei Shi², Quan Chen⁵, Xinzheng Du⁵, Li Yu⁵, Min-Jie Cao⁴, Zhulou Wang³, Shaohui Huang⁷, Tengchuan Jin⁸, Qiwei Wang⁹

Affiliations

¹ Division of Molecular Medicine, Hefei National Laboratory for Physical Sciences at Microscale, CAS Key Laboratory of Innate Immunity and Chronic Disease, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, China; Zhongshan Ophthalmic Center, Xian Lie South Road #54, Guangzhou, Guangdong, China.
² Department of Ophthalmology, The First Affiliated Hospital, University of Science and Technology of China, Hefei, Anhui, China.
³ Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.
⁴ College of Food and Biological Engineering, Jimei University, Xiamen, Fujian, China.
⁵ Division of Molecular Medicine, Hefei National Laboratory for Physical Sciences at Microscale, CAS Key Laboratory of Innate Immunity and Chronic Disease, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, China.
⁶ Zhongshan Ophthalmic Center, Xian Lie South Road #54, Guangzhou, Guangdong, China.
⁷ Institute of Biophysics, Chinese Academy of Sciences, Beijing, China; School of Biological Sciences, University of Chinese Academy of Sciences, Beijing, China.
⁸ Division of Molecular Medicine, Hefei National Laboratory for Physical Sciences at Microscale, CAS Key Laboratory of Innate Immunity and Chronic Disease, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, China; CAS Center for Excellence in Molecular Cell Science, Shanghai, China. Electronic address: jint@ustc.edu.cn.
⁹ Zhongshan Ophthalmic Center, Xian Lie South Road #54, Guangzhou, Guangdong, China. Electronic address: wang_qiwei@126.com.

PMID: 31254514
DOI: 10.1016/j.exer.2019.107712

Abstract

Crystallins are structural proteins in the lens that last a lifetime with little turnover. Deviant in crystallins can cause rare but severe visual impairment, namely, congenital cataracts. It is reported that several mutations in the acidic β-crystallin 4 (CRYBA4) are related to congenital cataracts. However, the pathogenesis of these mutants is not well understood at molecular level. Here we evaluate the biochemical properties of wild type CRYBA4 (CRYBA4^WT) and a pathogenic G64W mutant (CRYBA4^G64W) including protein folding, polymerization state and protein stability. Furthermore, we explore the differences in their interactions with α-crystallin A (CRYAA) and basic β-crystallin 1 (CRYBB1) via yeast two-hybrid and pull-down assay in vitro, through which we find that G64W mutation leads to protein misfolding, decreases protein stability, blocks its interaction with CRYBB1 but maintains its interaction with CRYAA. Our results deepen our understanding of the pathogenesis of congenital cataracts.

Keywords: Acidic β-crystallin 4; Congenital cataracts; Fluorescence correlation spectroscopy; G64W mutant; Protein folding; Protein interaction.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cataract* / congenital
Cataract* / genetics
Cataract* / metabolism
Humans
Lens, Crystalline / metabolism*
Mutation
Protein Folding*
beta-Crystallin A Chain / genetics*
beta-Crystallins / chemistry*

Substances

CRYBA4 protein, human
beta-Crystallin A Chain
beta-Crystallins