Britain's Andy Murray and Japan's Kei Nishikori will also be among those who drop their rackets and compete with their feet ahead of their opening matches in a $9 million ATP and WTA hardcourt event.
The tennis stars will face the Fort Lauderdale Strikers, a second-level US pro squad, at Ransom Everglades High School with a dinner afterward. Proceeds will be donated to the American Red Cross relief effort in Japan.
"I'm very concerned for the people of Japan and think it is only right that we do whatever we can to help those in need," Djokovic said. "I would love to see all of Miami support both events."
Serbia's Djokovic is off to an 18-0 start to the season, the best since Ivan Lendl's 25-0 start a quarter-century ago, and comes off an Indian Wells finals triumph over Spain's Nadal last Sunday.
Others ATP players set to take the field include Frenchman Richard Gasquet; Spaniards David Ferrer, Fernando Verdasco and Feliciano Lopez; Serbian Viktor Troicki, Austrian Jurgen Melzer and Marcos Baghdatis of Cyprus.