Journal Article

Inhibition of the mechano-enzymatic amyloidogenesis of transthyretin: role of ligand affinity, binding cooperativity and occupancy of the inner channel

Hawkins, Philip N., Verona, Guglielmo, Pepys, Mark B., Porcari, Riccardo, Raimondi, Sara, Bellotti, Vittorio, Faravelli, Giulia, Mangione, P. Patrizia, Giorgetti, Sofia, Taylor, Graham W., Corazza, Alessandra, Gillmore, Julian D.

2017

Overview

Dissociation of the native transthyretin (TTR) tetramer is widely accepted as the critical step in TTR amyloid fibrillogenesis. It is modelled by exposure of the protein to non-physiological low pH in vitro and is inhibited by small molecule compounds, such as the drug tafamidis. We have recently identified a new mechano-enzymatic pathway of TTR fibrillogenesis in vitro , catalysed by selective proteolytic cleavage, which produces a high yield of genuine amyloid fibrils. This pathway is efficiently inhibited only by ligands that occupy both binding sites in TTR. Tolcapone, which is bound with similar high affinity in both TTR binding sites without the usual negative cooperativity, is therefore of interest. Here we show that TTR fibrillogenesis by the mechano-enzymatic pathway is indeed more potently inhibited by tolcapone than by tafamidis but neither, even in large molar excess, completely prevents amyloid fibril formation. In contrast, mds84, the prototype of our previously reported bivalent ligand TTR ‘superstabiliser’ family, is notably more potent than the monovalent ligands and we show here that this apparently reflects the critical additional interactions of its linker within the TTR central channel. Our findings have major implications for therapeutic approaches in TTR amyloidosis.

• Publisher: Nature Publishing Group UK,Nature Portfolio
• Subject: 101/28 / 631/45/535 / 631/45/612 / 82 / 82/29 / 82/80 / 82/83 / Amyloid - metabolism / Benzophenones - pharmacology / Benzoxazoles - pharmacology / Binding Sites - drug effects / Fenamates - pharmacology / Humanities and Social Sciences / Humans / Models, Molecular / Molecular Structure / multidisciplinary / Nitrophenols - pharmacology / Prealbumin - chemistry / Prealbumin - drug effects / Prealbumin - metabolism / Protein Binding - drug effects / Protein Multimerization / Proteolysis / Science / Science (multidisciplinary) / Tolcapone