The foursome will compete in the TaylorMade Driving Relief - a two-team skins challenge over 18 holes - at Seminole golf course on May 17.
World number one Rory McIlroy will team up with Johnson and go head-to-head with Fowler and Wolff, with up to 3million (£2.38m) to American Nurses Foundation and CDC Foundation.
All four players will follow strict social distancing measures and comprehensive testing will be utilised to protect players, television production staff and others on the Florida site.
The challenge will be broadcast live simultaneously in the United States and the UK. "We are excited about the safe and responsible return of live golf and the opportunity to raise significant funds for those on the front lines of the COVID-19