title: Cosolvent effects on the fibrillation reaction of human IAPP.
abstract: Owing to the presence of various types of osmolytes in the cellular environment, this study focuses on the impact of stabilizing (TMAO and betaine) as well as destabilizing (urea) cosolvents on the aggregation and fibrillation reaction of the highly amyloidogenic islet amyloid polypeptide (IAPP). IAPP is associated with type-2 diabetes mellitus and is responsible for the disease accompanying β-cell membrane permeabilization and final β-cell loss. To reveal the impact of the cosolvents on the aggregation kinetics, conformational and morphological changes upon IAPP fibrillation, Thioflavin T fluorescence spectroscopy, atomic force microscopy and attenuated total reflection Fourier-transform infrared spectroscopy were applied. For TMAO, and less pronounced for betaine, a decrease of the growth rate of fibrils is observed, whereas the lag phase remains essentially unchanged, indicating the ability of the compatible solutes to stabilize large oligomeric and protofibrillar structures and therefore hamper fibril elongation. Conversely, urea displays concentration-dependent prolongation of the lag phase, indicating stabilization of IAPP in its unfolded monomeric state, hence leading to retardation of IAPP nuclei formation. Mixtures of urea with TMAO, and to a lesser extent with betaine, exhibit a counteractive effect. TMAO is able to fully compensate the prolonged lag phase induced by urea. This strongly matches the findings of a counteraction of TMAO and urea in protein folding and unfolding experiments. The data also reveal that the influence of these cosolvents is only on the aggregation kinetics without markedly changing the final IAPP fibrillar morphology, i.e., the solution structure and cosolvent composition essentially affect the kinetics of the fibrillation process only.
authors.affiliation: Physical Chemistry I-Biophysical Chemistry, Faculty of Chemistry, TU Dortmund University, Dortmund, Germany.
authors.forename: Janine
authors.initials: J
authors.lastname: Seeliger
authors.forename: Kathrin
authors.initials: K
authors.lastname: Estel
authors.forename: Nelli
authors.initials: N
authors.lastname: Erwin
authors.forename: Roland
authors.initials: R
authors.lastname: Winter
mesh_terms: D000682:Amyloid / Q000737:chemistry / Q000378:metabolism*; D001622:Betaine / Q000378:metabolism*; D003924:Diabetes Mellitus, Type 2 / Q000378:metabolism; D006801:Humans; D058228:Islet Amyloid Polypeptide / Q000737:chemistry / Q000378:metabolism*; D008744:Methylamines / Q000378:metabolism*; D055550:Protein Stability; D017433:Protein Structure, Secondary; D014508:Urea / Q000378:metabolism*
chemical_list: D000682:Amyloid; D058228:Islet Amyloid Polypeptide; D008744:Methylamines; D001622:Betaine; D014508:Urea; C005855:trimethyloxamine