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The chronic lead intoxication was modelled by injecting outbred rats intraperitoneally with lead acetate 3 times a week for 5 weeks. Using an in vitro motility assay, it was shown that lead intoxication causes changes in in vitro actin–myosin interaction, specifically, a decrease in the maximum sliding velocity of native thin filaments over myosin isolated from the left ventricular myocardium of the rat heart. No statistically significant changes were found in the calcium sensitivity and cooperativity of the “pCa–velocity” curve, characteristics of the motile filament fraction, and isometric force. Using electrophoretic separation of proteins, a shift in the ratio of myosin heavy chains toward β-chains with a lower ATPase activity was found.