The 41-year-old seven-time champion clocked a best time of one minute and 34.893 seconds to outpace his old rival, four-time champion Verstappen, by 0.168 seconds at the Circuit of the Americas.
Hamilton's Ferrari teammate Charles Leclerc was third after having several laps deleted, like many of the drivers, for exceeding track limits, finishing ahead of Antonelli and his Mercedes teammate and nearest title rival George Russell.
Antonelli leads Russell by 102 points in the championship and can become Formula 1's youngest champion, at 20, and Italy's first since Alberto Ascari of Ferrari in 1953, if he wins Sunday's race and his rivals fail to stay within reach of his points total.
Russell needs to finish in the top eight and Hamilton in the top four to keep their respective title hopes alive if Antonelli wins.
Hamilton's fastest lap ensured Ferrari had topped the times in all three practice sessions after Leclerc topped both on Friday. Leclerc was fast again in this session but had five laps deleted due to exceeding track limits, as a total of 42 laps were ruled out overall.