Formation of lipid-protein adducts in low-density lipoprotein by fluxes ofperoxynitrite and its inhibition by nitric oxide

Peroxynitrite (PN), the product of the diffusion-limited reaction between n itric oxide ((NO)-N-.) and superoxide (O-2(.-)), represents a relevant medi ator of oxidative modifications in low-density lipoprotein (LDL). This work shows for the first time the simultaneous action of low-controlled fluxes of PN and (NO)-N-. on LDL oxidation in terms of lipid and protein modificat ions as well as oxidized lipid-protein adduct formation. Fluxes of PN (e.g. ,1 muM min(-1)) initiated lipid oxidation in LDL as measured by conjugated dienes and cholesteryl ester hydroperoxides formation. Oxidized-LDL exhibit ed a characteristic fluorescent emission spectra (lambda (exc) = 365 nm, la mbda (max) = 417 nm) in parallel with changes in both the free amino groups content and the relative electrophoretic mobility of the particle. Physiol ogically relevant fluxes of (NO)-N-. (80-300 nM min(-1)) potently inhibited these PN-dependent oxidative processes. These results are consistent with PN-induced adduct formation between lipid oxidation products and free amino groups of LDL in a process prevented by the simultaneous presence of (NO)- N-.. The balance between rates of PN and (NO)-N-. production in the vascula r wall will critically determine the final extent of LDL oxidative modifica tions leading or not to scavenger receptor-mediated LDL uptake and foam cel l formation. (C) ?001 Academic Press.