Tottenham have agreed a partnership with the NFL to host two games annually at their new stadium for 10 years.
The NFL has staged regular season games at Wembley Stadium since 2007 and held three games at the ground last season.
There is optimism that the NFL will extend its agreement with Wembley beyond 2016, but the league has taken another step to further its international profile with the announcement of the deal with Tottenham on Wednesday.
Tottenham's new stadium, which will be built on the site of White Hart Lane, is due to open in 2018 and is to feature a retractable grass pitch with an artificial surface underneath for use in NFL games.
The NFL has also indicated that further games at other venues in the UK could be held in addition to the minimum of two at Tottenham's stadium during the 10-year agreement.
"With growing enthusiasm for the NFL in the United Kingdom, we are committed to hosting NFL games in world-class venues and are excited to partner with Tottenham Hotspur to play games at their future stadium," NFL commissioner Roger Goodell said.
"We share a vision and commitment to creating the best experience for our teams, fans and the local community."
London mayor Boris Johnson also praised the new agreement and talked up the prospect of an NFL franchise in the capital.
Johnson said: "Anyone who has seen American Football at Wembley Stadium cannot fail to have been thrilled by the spectacle.
"And the wise heads that run the NFL have clearly not missed the fact that Londoners are going absolutely gangbusters for Gridiron.
"Touchdowns in Tottenham can only add to our reputation as a global sporting powerhouse, and help us take another step towards our goal of having a permanent NFL franchise here in London."
Tottenham chairman Daniel Levy added: "We have an opportunity now to deliver one of the most unique sporting and entertainment venues in the world, bringing together the EPL [Premier League] and NFL for the first time."
