title: The mechanism of enhanced insulin amyloid fibril formation by NaCl is better explained by a conformational change model.
abstract: The high propensity of insulin to fibrillate causes severe biomedical and biotechnological complications. Insulin fibrillation studies attain significant importance considering the prevalence of diabetes and the requirement of functional insulin in each dose. Although studied since the early years of the 20(th) century, elucidation of the mechanism of insulin fibrillation has not been understood completely. We have previously, through several studies, shown that insulin hexamer dissociates into monomer that undergoes partial unfolding before converting into mature fibrils. In this study we have established that NaCl enhances insulin fibrillation mainly due to subtle structural changes and is not a mere salt effect. We have carried out studies both in the presence and absence of urea and Gdn.HCl and compared the relationship between conformation of insulin induced by urea and Gdn.HCl with respect to NaCl at both pH 7.4 (hexamer) and pH 2 (monomer). Fibril formation was followed with a Thioflavin T assay and structural changes were monitored by circular dichroism and size-exclusion chromatography. The results show salt-insulin interactions are difficult to classify as commonly accepted Debye-Hückel or Hofmeister series interactions but instead a strong correlation between the association states and conformational states of insulin and their propensity to fibrillate is evident.
authors: Department of Biological Chemistry and Life Sciences Institute, University of Michigan, Ann Arbor, Michigan, United States of America.
authors: Mahvish
authors: M
authors: Muzaffar
authors: Atta
authors: A
authors: Ahmad
mesh_terms: D000682:Amyloid / Q000737:chemistry*; D052160:Benzothiazoles; D002850:Chromatography, Gel; D002942:Circular Dichroism; D018360:Crystallography, X-Ray; D006801:Humans; D007328:Insulin / Q000737:chemistry*; D007700:Kinetics; D008958:Models, Molecular; D011487:Protein Conformation; D012965:Sodium Chloride / Q000494:pharmacology*; D013844:Thiazoles / Q000378:metabolism
chemical_list: D000682:Amyloid; D052160:Benzothiazoles; D007328:Insulin; D013844:Thiazoles; C009462:thioflavin T; D012965:Sodium Chloride