Wembley Stadium will stage a second NFL game in 2013 with the Minnesota Vikings to face the Pittsburgh Steelers on 29 September.
The London venue has hosted a regular-season game every year since 2007, but this is the first time there will be more than one fixture as the Jacksonville Jaguars take on San Francisco 49ers the following month on 27 October.
This additional match has been under discussion for some time and was approved by NFL owners at a meeting in Chicago.
"Since we started playing regular-season games in London five years ago, we have heard very clearly from our UK fans - they want more football," said NFL commissioner Roger Goodell.
"We are excited to play two games in London and take this next step in the growth of our game. We believe that more football will lead to more fans."
The Vikings will give up a home game in order to make the trip, with this being a one-year commitment from the Minneapolis-based franchise.
The Jaguars have committed to hosting games in London each year from 2013 to 2016 in a bid to establish a foothold in Britain.
"This is a unique opportunity for the Vikings organisation to highlight our brand on an international level," said Vikings owner and president Mark Wilf.
Steelers president Art Rooney II said: "We are thrilled that our players and Steelers fans around the world will be able to enjoy the atmosphere at historic Wembley Stadium."