Barcelona's Nou Camp will host rugby league for the first time next year, with Catalans Dragons facing Super League champions Wigan on 18 May.
Catalans, based 120 miles from Barcelona in the French city of Perpignan, were invited to the ground by the La Liga giants after winning rugby league's Challenge Cup in August.
Wigan chairman Ian Lenagan said it will be a "landmark moment" for the game.
"It will provide a shot in the arm for the new-era of Super League," he added.
Taking the match to the biggest stadium in Europe, with a capacity of 99,354, is a huge undertaking for the French rugby league club whose Stade Gilbert Brutus ground holds just 13,000. The population of Perpignan itself is around 121,000.
It will be the second time that Catalans will play in the Spanish city, having previously faced Warrington Wolves at the Olympic Stadium in 2009.
Dragons chairman Bernard Guasch said he hoped the match "is the beginning of a great story" between the French side and the Spanish giants.
Barcelona president Josep Maria Bartomeu hopes the rugby league game will match the interest generated when they hosted the final of French rugby union's Top 14 competition - a match that attracted a crowd of 99,000.
"I encourage the Dragons supporters to treat our stadium like their home and for Wigan and Barca fans to also share the experience of seeing rugby league played at the Camp Nou," Bartomeu said.
The Nou Camp is the fourth football stadium to host English rugby league matches in 2019, with the Challenge Cup remaining at Wembley, the Super League Grand Final at Old Trafford and Magic Weekend moving to Anfield.