Functional analyses and integrated mechanisms of cellular destruction by L-amino acid oxidase

Abstract

Snakebite accidents are prevalent worldwide and cause a spectrum of severe clinical manifestations and reduction of patient quality of life and economic income. L-amino acid oxidase (LAAO) is a highly toxic enzyme present in various venoms that causes tissue necrosis, oedema, coagulopathies, and organ failure. Here we investigate the mechanisms of LAAO cytotoxicity preceding cell death using recombinant LAAO and a catalytic inactive LAAO mutant. Wild-type LAAO uptake by cells leads to a decrease in lysosome number and size and inhibition of autophagy flux. Mitochondria function is impaired by severe proton leakage and mitochondrial fission is stimulated. Despite engulfment by autophagosomes, mitochondrial clearance is prevented by the lysosomal defects. The coordinate multi-organelle dysfunction strongly perturbs energy production, cell metabolism and clearance of defective organelles by autophagy, thereby triggering an irreversible destructive path. Considering the fast organelle impairment, strategies to reduce multi-organelle injury after LAAO exposure may be effective to maintain critical cell functions and strengthen adaptive responses against cytotoxicity.