Impact of hydrogen peroxide on structure, stability, and aggregational properties of human γS-crystallin

Cataract is the leading cause of blindness worldwide. Oxidative stress is one of the known risk factors for age-related cataracts. The present study was designed to understand the effect of H 2 O 2 -induced oxidative stress on human γS-crystallin and its relationship to lens opacification and catara...

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Published inJournal of biosciences Vol. 48; no. 1; p. 5
Main Author Vendra, Venkata Pulla Rao
Format Journal Article
LanguageEnglish
Published New Delhi Springer India 27.02.2023
Springer Nature B.V
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Summary:Cataract is the leading cause of blindness worldwide. Oxidative stress is one of the known risk factors for age-related cataracts. The present study was designed to understand the effect of H 2 O 2 -induced oxidative stress on human γS-crystallin and its relationship to lens opacification and cataract. Human γS-crystallin cDNA was cloned into the pET-20b vector, overexpressed in BL21 Star (DE3) cells, and was purified using ion-exchange and gel filtration chromatography. The structure, stability, and aggregational properties of human γS-crystallin under H 2 O 2 stress were studied using fluorescence and circular dichroism spectroscopy methods. H 2 O 2 treatment did not show any significant effect on the γS-crystallin secondary structure but showed an effect on its tertiary structure, resulting in N′-formylkynurenine formation. The H 2 O 2 -treated sample showed increased surface hydrophobicity, was less stable, and opened its Greek key motifs earlier with a midpoint of thermal unfolding curve (T m ) of 70.2°C compared with untreated γS-crystallin (T m =71.4°C). The sample treated with H 2 O 2 aggregated earlier in response to heating at 65°C. H 2 O 2 -induced oxidative stress alters the tryptophan microenvironment and the surface hydrophobicity of γS-crystallin, and these changes decrease its thermal stability and increase its tendency to aggregate, consistent with its role as a risk factor in age-related cataract.
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ISSN:0973-7138
0250-5991
0973-7138
DOI:10.1007/s12038-023-00330-w