Enzymatic and structural studies of human cardiac myosin from young and old subjects have been investigated to determine possible changes in myosin properties in aging hearts. Human ventricular myosin from old subjects (47-70 years old) has lower actin-activated ATPase activity than and increased alkaline sensitivity as compared to myosin from young subjects (1-132 months old). Ca2+-and K+(EDTA)-ATPase activities, pyrophosphate gel patterns and one-dimensional peptide mapping of heavy chains of ventricular myosin from old subjects are similar to those observed for myosin from young subjects. Atrial myosin from human hearts differs significantly from ventricular myosin in that the Ca2+-, Mg2+- and actin-activated myosin Mg2+-ATPase activities of atrial myosin are significantly higher than those of ventricular myosin. Pyrophosphate gel electrophoresis patterns and peptide mapping of heavy chains of atrial myosin are also different from those of ventricular myosin. Unlike ventricular myosin, atrial myosin from young hearts is similar to that of atrial myosin from old hearts in its enzymatic and structural properties.