Nitric oxide and peroxynitrite anion modulate liver plasma membrane fluidity and Na(+)/K(+)-ATPase activity

Abstract

Free radicals attack membranes and frequently alter their fluidity and function. The aim of the present work was to study the effect of nitric oxide (NO) radical and peroxynitrite anion on basolateral liver plasma membrane fluidity and on the activity of Na(+)/K(+)-ATPase. Basolateral membranes (BM) were isolated by ultracentrifugation in sucrose gradients and characterized enzymatically. BM were incubated with SNAP (a NO donor) or SIN-1 (a peroxynitrite donor). The release of NO or peroxynitrite was monitored by measuring NO(-)(2) + NO(-)(3). Relative fluidity was measured by polarization of fluorescence. NO increased membrane fluidity while peroxynitrite decreased it in a concentration-dependent manner. Na(+)/K(+)-ATPase activity was reduced by NO or peroxynitrite. Peroxynitrite anion inhibits ATPase activity in part by decreasing fluidity. However, it is very likely that both compounds inhibit ATPase activity by oxidation of the thiol groups of the enzyme. Our results suggest that NO may exert part of its biological effects by modulating membrane fluidity and function.