title: Solvent effects on the dynamics of amyloidogenic insulin revealed by neutron spin echo spectroscopy.
abstract: Insulin is well known to self-associate under specific solvent conditions. At low pH values, in the presence of sodium chloride (NaCl) and at elevated temperatures, insulin readily aggregates and forms amyloid fibrils. Without NaCl, but in the presence of ethanol, the lag time of this temperature-induced aggregation is increased drastically. In this study, we have analyzed the dynamical properties of bovine insulin under these two solvent conditions by using neutron spin echo (NSE) spectroscopy. In addition, small-angle X-ray scattering (SAXS) and thioflavin T (ThT) fluorescence experiments were carried out to track the concomitant structural changes of insulin. Measurements have mainly been performed at 318 K, where amyloid fibrils are formed over 25 h, when the insulin solution contains 100 mmol L(-1) of NaCl at pD = 2.4. In contrast, no amyloid fibrils are formed during 25 h at 318 K, when the insulin solution contains ethanol with a volume fraction of 20% at pD = 2.4. Remarkably, the NSE data reveal distinct dynamic signatures of insulin depending on the chosen solvent conditions. Collective diffusion of insulin molecules can be inferred from an increased diffusion coefficient at low wave vector transfers in the nonfibrillating sample, whereas self-diffusion is observed in the other case. The SAXS data confirm these dynamic behaviors because a pronounced correlation peak is only observed under conditions of collective diffusion. The dynamic responses of insulin, as revealed here by NSE spectroscopy, are in agreement with intermolecular interaction potentials derived recently from measurements of the static structure factors of insulin and lysozyme.
authors.affiliation: TU Dortmund University , Department of Chemistry and Chemical Biology, D-44221 Dortmund, Germany.
authors.forename: Mirko
authors.initials: M
authors.lastname: Erlkamp
authors.forename: Sebastian
authors.initials: S
authors.lastname: Grobelny
authors.forename: Antonio
authors.initials: A
authors.lastname: Faraone
authors.forename: Claus
authors.initials: C
authors.lastname: Czeslik
authors.forename: Roland
authors.initials: R
authors.lastname: Winter
mesh_terms: D000682:Amyloid / Q000737:chemistry*; D000818:Animals; D002417:Cattle; D007328:Insulin / Q000737:chemistry*; D007700:Kinetics; D033363:Neutron Diffraction / Q000379:methods*; D011487:Protein Conformation; D053838:Scattering, Small Angle; D012997:Solvents / Q000737:chemistry*; D013696:Temperature
chemical_list: D000682:Amyloid; D007328:Insulin; D012997:Solvents