Conversion of arginine into ornithine by advanced glycation in senescent human collagen and lens crystallins

Long lived proteins undergo age-related postsynthetic modifications that destabilize them by altering their conformation, charge, and helicity, thereby enhancing their resistance toward proteolysis and propensity to aggregate. The unexpected finding of substantial amounts of ornithine, the nonprotein amino acid, and decarbamidation product of arginine in acid hydrolysates of lens crystallins and skin collagen led us to investigate its source and mechanism of formation. In order to exclude ornithine formation as an artifact of acid hydrolysis, proteins were reductively alkylated with formaldehyde to convert ornithine to dimethyl-ornithine. The proteins were assayed for carboxymethyl-ornithine and glycated ornithine (furornithine) by liquid chromatography coupled to electrospray ionization mass spectrometry. Ornithine in acid hydrolysates of human lens and skin proteins increased from 1 to 15 nmol/mg protein from ages 10 to 90 years, whereas dimethyl-ornithine increased from 0.5 to 15 and from 0 to 5 nmol/mg protein, respectively. Carboxymethyl-ornithine and furornithine increased with age in lens and skin from similar to0 to 60 and 0 to 180 pmol/mg protein, respectively. In collagen, ornithine was elevated above levels of nondiabetic controls only when both diabetes and end stage renal disease were present. The age-related increase of these modifications provides evidence for substantial in vivo formation of ornithine in aging human tissue proteins. The mechanism of ornithine formation is not known, but data suggest that arginine-derived advanced glycation end products might serve as precursors for the in vivo conversion of ornithine from arginine.