1. The dye SYPRO orange binds to amylin amyloid fibrils but not pre‐fibrillar intermediates
- Author
-
Wong, Amy G and Raleigh, Daniel P
- Subjects
Biochemistry and Cell Biology ,Biological Sciences ,Brain Disorders ,Aging ,Dementia ,Diabetes ,Alzheimer's Disease including Alzheimer's Disease Related Dementias (AD/ADRD) ,Acquired Cognitive Impairment ,Neurodegenerative ,Alzheimer's Disease ,Neurosciences ,Aetiology ,2.1 Biological and endogenous factors ,Metabolic and endocrine ,Amyloid ,Benzothiazoles ,Fluorescent Dyes ,Humans ,Islet Amyloid Polypeptide ,Kinetics ,Thiazoles ,SYPRO-orange ,amyloid ,thioflavin-T ,islet amyloid polypeptide ,amylin ,Computation Theory and Mathematics ,Other Information and Computing Sciences ,Biophysics ,Biochemistry and cell biology ,Medicinal and biomolecular chemistry - Abstract
Amyloid deposition underlies a broad range of diseases including multiple neurodegenerative diseases, systemic amyloidosis and type-2 diabetes. Amyloid sensitive dyes, particularly thioflavin-T, are widely used to detect ex-vivo amyloid deposits, to monitor amyloid formation in vitro and to follow the kinetics of amyloid self-assembly. We show that the dye SYPRO-orange binds to amyloid fibrils formed by human amylin, the polypeptide responsible for islet amyloid formation in type-2 diabetes. No fluorescence enhancement is observed in the presence of pre-fibrillar species or in the presence of non-amyloidogenic rat amylin. The kinetics of human amylin amyloid formation can be monitored by SYPRO-orange fluorescence and match the time course determined with thioflavin-T assays. Thus, SYPRO-orange offers an alternative to thioflavin-T assays of amylin amyloid formation. The implications for the interpretation of SYPRO-orange-based assays of protein stability and protein-ligand interactions are discussed.
- Published
- 2016