Caloric Restriction Attenuates Dityrosine Cross-Linking of Cardiac and Skeletal Muscle Proteins in Aging Mice

Abstract

Oxidative damage, particularly to proteins, has been widely postulated to be a major causative factor in the loss of functional capacity during senescence. The nature of the various mechanisms that may contribute to protein oxidation is only partially understood. In this study, concentrations of two markers for oxidative damage,o,o′-dityrosine ando-tyrosine, were determined using stable isotope dilution gas chromatography–mass spectrometry in four tissues of the mouse, namely heart, skeletal muscle, brain, and liver, during youth (4 months old), adulthood (14 months old), and old (30 months old) age. A comparison was made between mice that had access to unlimited calories with those that were restricted to 60% of the caloric intake of thead libitumregimen. Caloric restriction of this magnitude extends the average and maximum life span of mice by ∼40%.In vitrostudies demonstrated thato,o′-dityrosine was generated selectively in proteins exposed to tyrosyl radical.o-Tyrosine increased in proteins oxidized with hydroxyl radical, which also resulted in a variable increase ino,o′-dityrosine. In mice fedad libitum, levels ofo,o′-dityrosine increased with age in cardiac and skeletal muscle but not in liver or brain. In contrast,o-tyrosine levels did not rise with age in any of the tissues examined. These results suggest that tyrosyl radical-induced protein oxidation increases selectively with age in skeletal muscle and heart. Caloric restriction prevented the increase ino,o′-dityrosine levels in cardiac and skeletal muscle but did not influenceo-tyrosine levels in any of the four tissues. This selective increase ino,o′-dityrosine levels and its prevention by a life-prolonging caloric restriction regimen raise the possibility that oxidation of muscle proteins by tyrosyl radical contributes to the deterioration of cardiac and skeletal muscle function with advancing age.