Luminescent imaging of insulin amyloid aggregation using a sensitive ruthenium-based probe in the red region
A sensitive and selective strategy to identify insulin fibrils remains a challenge for researchers in amyloid protein research. Thus, it is critical to detect, in vitro, the species generated during amyloid aggregation, particularly the fibrillar species. Here we demonstrate that the luminescent com...
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Veröffentlicht in: | Journal of inorganic biochemistry 2021-11, Vol.224, p.111585-111585, Article 111585 |
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Sprache: | eng |
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Zusammenfassung: | A sensitive and selective strategy to identify insulin fibrils remains a challenge for researchers in amyloid protein research. Thus, it is critical to detect, in vitro, the species generated during amyloid aggregation, particularly the fibrillar species. Here we demonstrate that the luminescent complex cis-[Ru(phen)2(3,4Apy)2]2+ (RuApy; phen = 1,10-phenanthroline; 3,4Apy = 3,4-diaminopyridine) is a rapid, low-cost alternative to in vitro detection of fibrillar insulin, using conventional optical techniques. The RuApy complex displays emission intensity enhancement at 655 nm when associated with insulin, which enables imaging of the conformational changes of the protein's self-aggregation. The complex shows high sensitivity to fibrillar insulin with a limit of detection of 0.85 μM and binding affinity of 12.40 ± 1.84 μM which is comparable to those of Thioflavin T and Congo red, with the advantage of minimizing background fluorescence, absorption of light by biomolecules, and light scattering from physiologic salts in the medium.
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•The RuApy (Apy = 3,4-diaminopyridine) complex maps the amyloid aggregation of bovine insulin.•RuApy monitors insulin aggregation using the complex's luminescence at 655 nm.•RuApy shows high sensitivity to fibrillar insulin with a limit of detection of 0.85 μM.•Binding stoichiometry of RuApy:fibrillar insulin was determined to be 1:2. |
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ISSN: | 0162-0134 1873-3344 |
DOI: | 10.1016/j.jinorgbio.2021.111585 |