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• W2745212934 abstract "Deficiency in insulin secretion and function that characterize type 2 diabetes often requires administration of extraneous insulin, leading to injection-site amyloidosis. Insulin aggregation at neutral pH is not well understood. Although oligomer formation is believed to play an important role, insulin oligomers have not been fully characterized yet. Here, we elucidate similarities and differences between in vitro insulin aggregation at acidic and neutral pH for a range of insulin concentrations (2.5-100 μM) by using kinetic thioflavin T fluorescence, circular dichroism, atomic force and electron microscopy imaging. Importantly, we characterize the size distribution of insulin oligomers at different assembly stages by the application of covalent cross-linking and gel electrophoresis. Our results show that at the earliest assembly stage, oligomers comprise up to 40% and 70% of soluble insulin at acidic and neutral pH, respectively. While the highest oligomer order increases with insulin concentration at acidic pH, the opposite tendency is observed at neutral pH, where oligomers up to heptamers are formed in 10 μM insulin. These findings suggest that oligomers may be on- and off-pathway assemblies for insulin at acidic and neutral pH, respectively. Agitation, which is required to induce insulin aggregation at neutral pH, is shown to increase fibril formation rate and fibrillar mass both by an order of magnitude. Insulin incubated under agitated conditions at neutral pH rapidly aggregates into large micrometer-sized aggregates, which may be of physiological relevance and provides insight into injection-site amyloidosis and toxic pulmonary aggregates induced by administration of extraneous insulin." @default.
• W2745212934 created "2017-08-17" @default.
• W2745212934 creator A5041897244 @default.
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• W2745212934 date "2017-08-28" @default.
• W2745212934 modified "2023-10-01" @default.
• W2745212934 title "Elucidation of insulin assembly at acidic and neutral pH: Characterization of low molecular weight oligomers" @default.
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• W2745212934 doi "https://doi.org/10.1002/prot.25365" @default.
• W2745212934 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/28796342" @default.
• W2745212934 hasPublicationYear "2017" @default.
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