The 22-year-old missed Monday's training session with Roy Hodgson's and will sit out of both fixtures against Republic of Ireland and Brazil.
The Football Association confirmed the 22-year-old fullback arrived at St. George's Park on Sunday with an injury and has been subsequently ruled out after being assessed by England's medical staff.
Walker is believed to have picked up the injury during Tottenham's goalless friendly draw against against Jamaica in the Bahamas last Thursday. The FA also confirmed no replacement had been called up in Walker's absence.
Manchester United forward Danny Welbeck also missed Monday's training session, but the 22-year-old has not been ruled out of England's fixtures against the Republic of Ireland on May 29 and Brazil on June 2.