title: Structure and dynamics of a partially folded protein are decoupled from its mechanism of aggregation.
abstract: A common strategy to study the mechanism of amyloid formation is the characterization of the structure and dynamics of the precursor state, which is in most cases a partially folded protein. Here we investigated the highly dynamic conformational state formed by the protein domain HypF-N at low pH, before aggregation, using fluorescence, circular dichroism, and NMR spectroscopies. The NMR analysis allowed us, in particular, to identify the regions of the sequence that form hydrophobic interactions and adopt an alpha-helical secondary structure in the pH-denatured ensemble. To understand the role that this residual structure plays in the aggregation of this protein, we probed the mechanism of aggregation using protein engineering experiments and thus identified the regions of the sequence of HypF-N that play a critical role in the conversion of this dynamic state into thioflavin T-binding and beta-sheet containing protofibrils. The combination of these two complementary approaches revealed that the aggregation of pH-denatured HypF-N is not structure-dependent, meaning that it is not driven by the regions of the protein that are either less or more protected in the initial partially folded state. It is, by contrast, promoted by discrete protein regions that have the highest intrinsic propensity to aggregate because of their physicochemical properties.
authors.affiliation: Dipartimento di Scienze Biochimiche, Università di Firenze, Viale Morgagni 50, 50134 Firenze, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Calloni
authors.forename: Christofer
authors.initials: C
authors.lastname: Lendel
authors.forename: Silvia
authors.initials: S
authors.lastname: Campioni
authors.forename: Silva
authors.initials: S
authors.lastname: Giannini
authors.forename: Alessandra
authors.initials: A
authors.lastname: Gliozzi
authors.forename: Annalisa
authors.initials: A
authors.lastname: Relini
authors.forename: Michele
authors.initials: M
authors.lastname: Vendruscolo
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Dobson
authors.forename: Xavier
authors.initials: X
authors.lastname: Salvatella
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Chiti