She made her Broadway debut in the 1980 revival of West Side Story, and went on to appear in the 1983 film Scarface as Al Pacino's character's sister, Gina Montana, which proved to be her breakout role.

[1][2] Mastrantonio first appeared on screen in Brian De Palma's Scarface (1983) as Gina, sister of Al Pacino's Tony Montana.

She achieved prominence for her Oscar and Golden Globe-nominated role in The Color of Money (1986) opposite Paul Newman and Tom Cruise.

Mastrantonio has appeared on Broadway in various musicals, including West Side Story, Copperfield, The Human Comedy, and the 2002 revival of Man of La Mancha, where she played Aldonza/Dulcinea opposite Brian Stokes Mitchell.

In 1984, she was featured in a benefit performance of A Christmas Carol with Helen Hayes, Raul Julia, Harold Scott, F. MacIntyre Dixon, and Len Cariou at the Symphony Space in New York.

Starting with the final episode of Season 1 (2012), she had a recurring role on Grimm, playing Kelly Burkhardt, mother of the show's protagonist.

In 2015 she was cast in a leading role in Limitless on CBS, playing FBI Special Agent in Charge Nasreen "Naz" Pouran, which debuted on September 22, 2015.