Kinetic analysis of the multistep aggregation pathway of human transthyretin.

ABSTRACT: Aggregation of transthyretin (TTR) is the causative agent for TTR cardiomyopathy and polyneuropathy amyloidoses. Aggregation is initiated by dissociation of the TTR tetramer into a monomeric intermediate, which self-assembles into amyloid. The coupled multiple-step equilibria and low-concentration, aggregation-prone intermediates are challenging to probe using conventional assays. We report a ¹⁹F-NMR assay that leverages a highly sensitive trifluoroacetyl probe at a strategic site that gives distinct ¹⁹F chemical shifts for the TTR tetramer and monomeric intermediate and enables direct quantification of their populations during the aggregation process. Integration of real-time ¹⁹F-NMR and turbidity measurements as a function of temperature allows kinetic an

SUBMITTER: Sun X 

PROVIDER: S-EPMC6142231 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature