High-resolution ex vivo NMR spectroscopy of human Z α1-antitrypsin
Alistair M. Jagger, Christopher A. Waudby, James A. Irving, John Christodoulou, David A. Lomas
Abstract
Abstract Genetic mutations predispose the serine protease inhibitor α 1 -antitrypsin to misfolding and polymerisation within hepatocytes, causing liver disease and chronic obstructive pulmonary disease. This misfolding occurs via a transiently populated intermediate state, but our structural understanding of this process is limited by the instability of recombinant α 1 -antitrypsin variants in solution. Here we apply NMR spectroscopy to patient-derived samples of α 1 -antitrypsin at natural isotopic abundance to investigate the consequences of disease-causing mutations, and observe widespread chemical shift perturbations for methyl groups in Z AAT (E342K). By comparison with perturbations induced by binding of a small-molecule inhibitor of misfolding we conclude that they arise from rapid exchange between the native conformation and a well-populated intermediate state. The observation that this intermediate is stabilised by inhibitor binding suggests a paradoxical approach to the targeted treatment of protein misfolding disorders, wherein the stabilisation of disease-associated states provides selectivity while inhibiting further transitions along misfolding pathways.