Receptor-mediated toxicity of human amylin fragment aggregated by short- and long-term incubations with copper ions
| Autor: | V. G. Nicoletti, Giuseppe Caruso, Susan M. Lunte, Donatella A. Distefano, Claudia G. Fresta, Giuseppe Lazzarino, Paolo Parlascino |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Amyloid Cell viability Clinical Biochemistry Receptor for Advanced Glycation End Products Amylin Nerve Tissue Proteins Receptors Nerve Growth Factor Receptor-mediated cell toxicity Fibril RAGE (receptor) Cell Line 03 medical and health sciences Aggregation 0302 clinical medicine Glycation Animals Humans Viability assay Receptor Molecular Biology Copper Oligomers Chemistry Cell Biology General Medicine Islet Amyloid Polypeptide Rats 030104 developmental biology Biochemistry Toxicity 030217 neurology & neurosurgery |
| Popis: | Human amylin (hA1-37) is a polypeptide hormone secreted in conjunction with insulin from the pancreatic β-cells involved in the pathogenesis of type 2 diabetes mellitus (T2DM). The shorter fragment hA17-29 than full-length peptide is capable to form amyloids "in vitro". Here, we monitored the time course of hA17-29 β-amyloid fibril and oligomer formation [without and with copper(II)], cellular toxicity of different amyloid aggregates, and involvement of specific receptors (receptor for advanced glycation end-products, RAGE; low-affinity nerve growth factor receptor, p75-NGFR) in aggregate toxicity. Fibril and oligomer formation of hA17-29 incubated at 37 °C for 0, 48, and 120 h, without or with copper(II), were measured by the thioflavin T fluorescence assay and ELISA, respectively. Toxicity of hA17-29 aggregates and effects of anti-RAGE and anti-p75-NGFR antibodies were evaluated on neuroblastoma SH-SY5Y viability. Fluorescence assay of hA17-29 indicates an initial slow rate of soluble fibril formation (48 h), followed by a slower rate of insoluble aggregate formation (120 h). The highest quantity of oligomers was recorded when hA17-29 was pre-aggregated for 48 h in the presence of copper(II) showing also the maximal cell toxicity (-44% of cell viability, p |
| Databáze: | OpenAIRE |
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