Two months short of her 48th birthday, the holder of 58 grand slam titles will make her Olympic debut in the women's doubles with Lisa Raymond in the Athens tennis tournament which takes place from August 15 to 22.
Defending champion Venus Williams and younger sister Serena will compete in the women's singles along with Jennifer Capriati and Chanda Rubin, the United States Tennis Association said in a statement.
Venus and Serena will also attempt to retain the women's doubles gold they won in Sydney in 2000.
The men's singles team comprises US Open champion Andy Roddick, Mardy Fish, Vince Spadea and Taylor Dent. Roddick and Fish, together with twin brothers Bob and Mike Bryan, will also contest the doubles.
The 1996 Olympic men's champion, Andre Agassi, was ineligible as he has not made himself available to play Davis Cup tennis for the United States in the past two years.
Navratilova is eligible as she played doubles in the US women's Fed Cup final defeat by France last November and also in their first round win over Slovenia in April.
The Czech-born American's appearance in Athens will extend a year in which she returned to grand slam singles action for the first time in a decade.
She lost in the first round at the French Open in May but the nine-times Wimbledon singles champion reached the second round at the All England Club this week before losing again to her conqueror in Paris, Gisela Dulko of Argentina.
SUCCESSIVE GOLDS
Venus, knocked out of Wimbledon in a second round upset on Thursday, is attempting to become the first player to win successive Olympic golds in singles.
American women have won the last three Olympic singles titles.
Capriati, 28, won the women's singles gold at the age of 16 in Barcelona in 1992, while Lindsay Davenport triumphed in Atlanta four years later.
"As a former Olympian, I know that our team will experience one of the most memorable moments of their careers in Athens," said women's team captain Zena Garrison, who lost to Navratilova in the 1990 Wimbledon final.
Men's captain Patrick McEnroe said: "This team is excited and honoured to be participating in the Olympic Games and consider it a downright privilege.
"We are going to put forth the best possible effort to bring as many medals as we can back to the United States."
The Games themselves run from August 13 to August 29.
