R&B singer Keys has teamed up with the star of rock bands White Stripes and The Raconteurs for Another Way to Die, the first duet in Bond theme history.
The announcement follows speculation that the job would go to British stars Amy Winehouse, Duffy or Leona Lewis.
Quantum of Solace, which will see Daniel Craig return as 007, will be released later this year.
Winehouse was working on a theme with producer Mark Ronson - but that was scrapped because Ronson said the singer was not up to recording.
The new partnership sees the slick and soulful Keys team up with raw and bluesy rocker White.
Despite their different musical backgrounds, both are leading figures in the US scene, with 18 Grammy Awards between them.
White has written, produced and played drums on the track.
The film's producers Michael G Wilson and Barbara Broccoli said: "We are delighted and pleased to have two such exciting artists as Jack and Alicia."
The musicians "were inspired by our film to join together their extraordinary talents in creating a unique sound for Quantum of Solace", the producers said.
Previous iconic James Bond signature tunes have been recorded by stars including Dame Shirley Bassey, Duran Duran and Sir Paul McCartney.
The theme to the last film, 2006's Casino Royale, was recorded by another US rocker, Chris Cornell.
Quantum of Solace, the 22nd official film in the 007 series, picks up from Casino Royale as the secret agent sets out to avenge the apparent death of his lover.
It has a bigger budget than the last film and producers have promised more action.
The title is taken from a short story published by Bond creator Ian Fleming in 1960.
It will hit cinemas on 31 October in the UK, with the rest of the world seeing it from early November.
David Arnold will compose the film's score, with the soundtrack to be released on 28 October.