Summary & Purpose

Purpose: The significant lens chaperone protein α-crystallin (αABc) comprising αA-crystallin (αAc) and αB-crystallin (αBc) subunits is found to form membrane-bound aggregates with age and cataract formation. However, the molecular basis for such aggregate formation is still unclear. Our research aims to elucidate the effect of lipids (phospholipids and sphingolipids) and cholesterol (Chol) peroxidation on the aggregation of αAc, αBc, and αABc to bovine lens nuclear membrane (NM).

Methods: Lipid and Chol peroxidation were induced by photosensitized peroxidation reaction, and topographical images of NM and oxidized-NM (Ox-NM) with and without incubation with αAc, αBc, or αABc were obtained using atomic force microscopy (AFM).

Results: The percentage of membrane area occupied (PMAO) on NM by aggregation of αAc, αBc, or αABc with NM was significantly smaller without lipid and Chol peroxidation in NM. However, with lipid and Chol peroxidation in NM, the PMAO on Ox-NM by aggregation of αAc, αBc, or αABc with Ox-NM was significantly more extensive, and PMAO increases with an increase in lipid and Chol peroxidation. Large-size aggregates of αAc, αBc, or αABc on Ox-NM with the depressed central region for αAc and αABc aggregates on Ox-NM were observed at the greater extent of lipid and Chol peroxidation.

Conclusions: Lipid and Chol peroxidation induce membrane defects on NM followed by extensive aggregation of αAc, αBc, and αABc on Ox-NM, suggesting that lipid and Chol peroxidation promotes aggregation of αAc, αBc, and αABc to Ox-NM and formation of such large aggregates likely promote increased light scattering and cataract formation.

Author Identifier

Nawal K. Khadka, ORCID: 0000-0002-2404-3799

Preston Hazen, ORCID: 0000-0002-1713-3980

Xinzhu Pu, ORCID: 0000-0002-5780-1664

Laxman Mainali, ORCID: 0000-0002-9570-9041

Date of Publication or Submission

9-16-2025

DOI

https://doi.org/10.18122/biophysics_data.7.boisestate

Funding Citation

Supported by a grant from the National Eye Institute and National Institute of General Medical Sciences of the National Institutes of Health (R01EY030067) via Institutional Development Award co-funding, and by the Biomolecular Research Core at Boise State University (RRID:SCR_019174) through a grant from the National Science Foundation (2320410). The authors thank the Surface Science Laboratory at Boise State University for providing an atomic force microscopy facility. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

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Series

Data Format

*.xlsx, *.spm, *.raw, and *.txt

File Size

1.27 GB

Data Attributes

See README.txt

Time Period

2023-2025

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Comments

Dataset Update 09/18/2025: Thermo.raw files for LC-HRMS data were added for Fig. 3.

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