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Police said 61-year-old Ronnie died at 0907 GMT after being transferred to Wexham Park hospital, Slough, from a hospital in Ascot.
The man, once part of the infamous gang "The Firm", is understood to have suffered a heart attack.
Ronnie, a homosexual who had been married, was taken to Heatherwood Hospital, Ascot, two days ago after collapsing in his room at Broadmoor.
Reign of terror
He was transferred to Wexham Park hospital last night after his condition deteriorated, and he later died there.
Kray and his twin brother Reggie were sentenced to 30 years in prison in 1969, which ended a 10 year bloody reign of terror in London.
Ronnie had shot George Cornell in the Blind Beggar public house in Whitechapel in 1966 for calling him a "fat poof".
And a year later Reggie stabbed Jack "The Hat" McVitie in a flat in North London.
Ronnie was later judged to be criminally insane and sent to the Broadmoor secure hospital.
He told friends and family he expected to die a prisoner.
Kray, who shared the ward with Yorkshire Ripper Peter Sutcliffe, had suffered two earlier heart attacks, the latest in September 1993 after which doctors warned his rumoured 100 cigarettes a day habit would kill him.
The Krays have reached iconic status, revered by some and scorned by others.
They ran a brutal gang in London's East End during the late 1950s and 1960s which netted them a fortune and allowed them to live a life of luxury.
Since their conviction, an industry has grown around them with books, T-shirts, television specials and a film starring pop star twins Gary and Martin Kemp of Spandau Ballet.
Reggie is understood to have learnt of his twin's death from a fellow prisoner in Maidstone jail who had heard it on the radio and was described as "absolutely distraught".
Their elder brother Charlie, who served seven years for his part in the crimes, said he was saddened by the loss and that his late brother had been misunderstood.
Scotland Yard had been on the trail of the Krays for many years and finally caught up with them and their accomplices in 1968.
In June 1997 Charlie was found guilty of masterminding a £39m cocaine plot and jailed for 12 years. He died in hospital as an inmate in April 2000.
The surviving brother Reggie, who had hoped for parole after serving 30 years in prison, got release as a dying wish when Home Secretary Jack Straw ordered his freedom when he was diagnosed with terminal cancer and given just weeks to live.
He died in October 2000.
Neither the Krays' incarceration or death has suppressed their legend.
Many people had campaigned for their release as they were seen as having a sense of honour by ridding London's streets of criminals.
Despite using violence many insisted women and children were safe as long as the Krays prowled the streets.
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