Three years on from suffering penalty shootout heartache to Italy in the delayed Euro 2020 tournament, England were once again forced to accept the runners-up prize at the Olympiastadion in Berlin.
Southgate's men were dominated from the first whistle but managed to hold out until half time, where they received a major boost in the form of Rodri being taken off with an injury.
However, La Roja found a sixth gear in attack at the start of the second period, culminating in Lamine Yamal teeing up Nico Williams to clinically fire in the first goal of the showpiece match.
England were at sixes and sevens at the back all of a sudden, but Luis de la Fuente's men failed to add to their advantage and were pegged back by Three Lions super sub Cole Palmer, who produced a fabulous finish from outside of the box.
King Charles sends 'heartfelt sympathies' to England
Palmer's strike appeared to be just what the doctor ordered to kick Spain back into life again, though, and with just four minutes of normal time remaining, Marc Cucurella's inch-perfect pass for Mikel Oyarzabal ended with the Real Sociedad man tapping in the clincher.
As Spain celebrated becoming European champions for an unrivalled fourth time, England's inconsolable troops were left to rue what could have been again, as they became the first nation to lose back-to-back Euros finals.
The Prince of Wales and president of the Football Association - Prince William - attended the Berlin final alongside his eldest child Prince George of Wales, as was the case in the Euro 2020 final at Wembley.
King Charles III directly addressed Gareth Southgate after England's crushing loss, expressing his "heartfelt" sympathy towards him and his players and insisting that "many triumphs" lie ahead for them.
"Although victory may have eluded you this evening, nevertheless my wife and I join all my family in urging you and your support team to hold your heads high," a statement from Buckingham Palace read.
"All those who have participated in sporting activities at any level will know how utterly despairing such a result can feel when the prize was so near - and will join me in sending heartfelt sympathy, even as we congratulate Spain.
"But please know that your success in reaching the European Championship final is a really great achievement in itself, and one that brings with it the pride of a nation which will continue to roar for the Three Lions today - and in the many triumphs which I have no doubt lie ahead. Charles R".
Speaking to broadcasters after the final whistle, captain Harry Kane - who was taken off just after the hour mark following a frustrating evening - admitted that the loss would hurt for "a long time".
Meanwhile, Southgate - whose future with the England team has been left shrouded in even more uncertainty - conceded that he was "devastated for everyone" before batting away questions about whether Sunday's game could have been his last with the Three Lions.