1. Label-free visualization of unfolding and crosslinking mediated protein aggregation in nonenzymatically glycated proteins.
- Author
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Mukunda, Darshan Chikkanayakanahalli, Basha, Shaik, D'Souza, Meagan Gail, Chandra, Subhash, Ameera, K., Stanley, Weena, Mazumder, Nirmal, and Mahato, Krishna Kishore
- Subjects
PROTEIN crosslinking ,ADVANCED glycation end-products ,DENATURATION of proteins ,SOIL structure ,BIOFLUORESCENCE ,RECEPTOR for advanced glycation end products (RAGE) ,GLYOXALASE - Abstract
Nonenzymatic glycation (NEG) unfolds and crosslinks proteins, resulting in aggregation. Label-free evaluation of such structural changes, without disturbing molecular integrity, would be beneficial for understanding the fundamental mechanisms of protein aggregation. The current study demonstrates the assessment of NEG-induced protein aggregation by combining autofluorescence (AF) spectroscopy and imaging. The methylglyoxal (MG) induced protein unfolding and the formation of cross-linking advanced glycation end-products (AGEs) leading to aggregation were evaluated using deep-UV-induced-autofluorescence (dUV-AF) spectroscopy in proteins with distinct structural characteristics. Since the AGEs formed on proteins are fluorescent, the study demonstrated the possibility of autofluorescence imaging of NEG-induced protein aggregates. Autofluorescence spectroscopy can potentially reveal molecular alterations such as protein unfolding and cross-linking. In contrast, AGE-based autofluorescence imaging offers a means to visually explore the structural arrangement of aggregates, regardless of whether they are amyloid or non-amyloid in nature. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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