With Serena and Venus Williams unavailable together with Sloane Stephens and Madison Keys, captain Kathy Rinaldi's top-ranked player is world number 35 Danielle Collins.
She will be joined in Prague by 48th ranked Sofia Kenin, number 63 Alison Riske and Nicole Melichar, the world's 15th ranked doubles player.
The tie takes place on Nov 10-11.
Riske was a member of the U.S. squad that won the Fed Cup title over Belarus last year in Minsk but this is a first Fed Cup nomination for Collins, Kenin and Melichar.
Stephens is the highest American on the WTA rankings at sixth while Keys is 16th. The Williams sisters Serena and Venus are currently ranked 15th and 40th.
The Czechs will be rolling out their best to face the U.S. with world number seven Petra Kvitova and number eight Karolina Pliskova along with doubles number one Katerina Siniakova and number five Barbora Strycova. All are expected to see action in the best-of-five match series inside Prague's O2 Arena.
The U.S. have dominated the Czechs in Fed Cup play posting a 10-2 record. The nations have met twice in the final -- with Czechoslovakia winning in 1985 and the U.S. winning in 1986.