H101G Mutation in Rat Lens αB-Crystallin Alters Chaperone Activity and
Divalent Metal Ion Binding

Yi-Ying      Wu; Naveen   Kumar Reddy    Desu; Shou-Yun      Lu; Bi-Yu      Yu; Ramya      Kumar; Fu-Yung       Huang

doi:10.2174/1389201022666210702130843

Abstract

Background: The molecular chaperone function of αB-crystallins is heavily involved in maintaining lens transparency and the development of cataracts.

Objectives: The aim of the study was to investigate whether divalent metal ion binding improves the stability and αB-crystallin chaperone activity.

Methods: In this study, we have developed an H101G αB-crystallin mutant and compared the surface hydrophobicity, chaperone activity, and secondary and tertiary structure with the wild type in the presence and absence of metal ions.

Results: Substitution of His101 with glycine resulted in structural and functional changes. Spectral analysis and chaperone-like activity assays showed that substitution of glycine resulted in a higher percentage of random coils, increased hydrophobicity, and 22±2% higher chaperone-like activity. Whereas in the presence of the Cu2+ ion, H101G exhibited 32±1% less chaperone-like activity compared to the wild type.

Conclusion: Cu2+ has been reported to enhance the chaperone-like activity of lens α-crystallin. Our results indicate that H101 is the predominant Cu2+ binding site, and the mutation resulted in a partial unfolding that impaired the binding of Cu2+ to H101 residue. In conclusion, this study further helps to understand the important binding site for Cu2+ to αB-crystallin.

Keywords: αB-crystallin, chaperone activity, hydrophobicity, site-directed mutagenesis, circular dichroism, far-UV CD spectra.

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DOI https://dx.doi.org/10.2174/1389201022666210702130843	Print ISSN 1389-2010
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H101G Mutation in Rat Lens αB-Crystallin Alters Chaperone Activity and Divalent Metal Ion Binding

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