Determination of Dideoxyosone Precursors of AGEs in Human Lens Proteins
| Autor: | Monika Pischetsrieder, Trilok J. Parmar, S.R. Kaid Johar, Ram H. Nagaraj, Smitha Padmanabha, Mikhail Linetsky, Abhay R. Vasavada, Jasmin Meltretter |
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| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Adult
Glycation End Products Advanced Glycosylation Blotting Western Molecular Sequence Data Biophysics Enzyme-Linked Immunosorbent Assay Biochemistry Article Cataract Mass Spectrometry Lens protein chemistry.chemical_compound Young Adult Western blot Crystallin Glycation Protein purification Lens Crystalline medicine Animals Humans Amino Acid Sequence Pentosidine Molecular Biology medicine.diagnostic_test Lysine Reproducibility of Results Crystallins chemistry Cattle Glucosepane |
| Popis: | Dideoxyosones (DDOs) are intermediates in the synthesis of advanced glycation end products (AGEs), such as pentosidine and glucosepane. Although the formation of pentosidine and glucosepane in the human lens has been firmly established, the formation of DDOs has not been demonstrated. The aim of this study was to develop a reliable method to detect DDOs in lens proteins. A specific DDO trapping agent, biotinyl-diaminobenzene (3,4-diamino-N-(3-{[5-(2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanoyl]aminopropyl) benzamide) (BDAB) was added during in vitro protein glycation or during protein extraction from human lenses. In vitro glycated human lens protein showed strong reaction in monomeric and polymeric crosslinked proteins by western blot and ELISA. Glycation of BSA in the presence of BDAB resulted in covalent binding of BDAB to the protein and inhibited pentosidine formation. Mass spectrometric analysis of lysozyme glycated in the presence of BDAB showed the presence of quinoxalines at lysine residues at positions K1, K33, K96, and K116. The ELISA results indicated that cataractous lens proteins contain significantly higher levels of DDO than non-cataractous lenses (101.9±67.8 AU/mg protein vs. 31.7±19.5 AU/mg protein, p |
| Databáze: | OpenAIRE |
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