In 2000, she starred with Harrison Ford in one of the biggest box office hits of the year, Robert Zemeckis's supernatural thriller What Lies Beneath. In 2001, she starred opposite Sean Penn in I Am Sam and in 2002 alongside Renée Zellweger and Robin Wright Penn in White Oleander, which earned her a Screen Actors Guild Award nomination as Best Supporting Actress.

She then took a lengthy hiatus from the spotlight but returned to acting in 2006. Her first projects teamed her with director Amy Heckerling in the romantic comedy I Could Never Be Your Woman (2007, opposite Paul Rudd), as well as opposite Robert De Niro, Claire Danes, and Sienna Miller in the fantasy epic Stardust. Pfeiffer co-starred in the film adaptation of the hit Broadway musical Hairspray as Velma Von Tussle. She will make a cameo appearance in The Prince and the Pauper, a film in which her sister Dedee stars. Pfeiffer received her star on the Hollywood Walk of Fame on August 6, 2007, in front of the Hollywood