Conceived as a full remake of the original, Mafia: Definitive Edition was built from the ground up with new assets and an expanded story, although missions and arcs from the original game are carried over. As with the 2002 game, players control Tommy Angelo throughout the game's single-player campaign, and its world is navigated on foot or by vehicle. New to the remake is the introduction of motorcycles, a first in the series. Mafia: Definitive Edition's gameplay mechanics are based on those of Mafia III. A 'Classic Difficulty' setting is also included, serving as the game's hardest difficulty setting that changes some gameplay mechanics such as ammunition management and police response to crimes, bringing the game closer to the original 2002 game.

Outside the main story mode, a separate Free Ride mode is included "as a nod to the original game", allowing the player to explore the city at their own leisure without any mandatory mission objectives. Unlike the original game, however, Free Ride and Free Ride Extreme are merged into one game mode, with the latter's outlandish and over-the-top side missions incorporated into the main Free Ride segment as hidden secrets to be uncovered by the player[

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