title: Time-resolved FRET detection of subtle temperature-induced conformational biases in ensembles of α-synuclein molecules.
abstract: The α-synuclein (αS) molecule, a polypeptide of 140 residues, is an intrinsically disordered protein that is involved in the onset of Parkinson's disease. We applied time-resolved excitation energy transfer measurements in search of specific deviations from the disordered state in segments of the αS backbone that might be involved in the initiation of aggregation. Since at higher temperatures, the αS molecule undergoes accelerated aggregation, we studied the temperature dependence of the distributions of intramolecular segmental end-to-end distances and their fast fluctuations in eight labeled chain segments of the αS molecule. Over the temperature range of 5-40 °C, no temperature-induced unfolding or folding was detected at the N-terminal domain (residues 1-66) of the αS molecule. The intramolecular diffusion coefficient of the segments' ends relative to each other increased monotonously with temperature. A common very high upper limiting value of ∼25 A²/ns was reached at 40 °C, another indication of a fully disordered state. Three exceptions were two segments with reduced values of the diffusion coefficients (the shortest segment where the excluded volume effect is dominant and the segment labeled in the NAC domain) and a nonlinear cooperative transition in the N-terminal segment. These specific subtle deviations from the common pattern of temperature dependence reflect specific structural constraints that could be critical in controlling the stability of the soluble monomer, or for its aggregation. Such very weak effects might be dominant in determination of the fate of ensembles of disordered polypeptides either to folding or to misfolding.
authors.affiliation: The Goodman Faculty of Life Sciences, Bar-Ilan University, 52900 Ramat Gan, Israel.
authors.forename: Asaf
authors.initials: A
authors.lastname: Grupi
authors.forename: Elisha
authors.initials: E
authors.lastname: Haas
mesh_terms: D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D046912:Multiprotein Complexes / Q000737:chemistry*; D011487:Protein Conformation; D013696:Temperature; D013997:Time Factors; D051844:alpha-Synuclein / Q000737:chemistry*
chemical_list: D046912:Multiprotein Complexes; D051844:alpha-Synuclein