Serena Williams and Venus Williams fell 7-6 (5) 6-4 to the Czech duo of Lucie Hradecka and Linda Noskova during the prime-time first round doubles match at the U.S. Open, which could be Serena's last.

Winners of 14 Grand Slam doubles titles, including two U.S. Opens, the Williams sisters were playing together for the first time in more than four years. They were handed a wild card into the draw for this year's U.S. Open just weeks after Serena indicated she planned to retire following the tournament.

The occasion attracted a capacity crowd to watch one of tennis's most successful partnerships take perhaps a final bow.

The Williams sisters were unable to take advantage of two opportunities to gain a crucial service break in the first set, and paid the price as Hradecka and Noskova clinched the tie break. 

Buoyed by their opening set performance, the Czechs scored an early break in the second and were soon 4-1 ahead with victory beckoning. 

The Williams' were determined not to go down without a fight however, and after holding serve dug out a service break and held again to level at 4-4. 

But hopes of an epic comeback were snuffed out rapidly with Noskova comfortably holding serve for a 5-4 lead. 

The Czech duo then took three match points at 0-40 on Serena's serve, converting on the second opportunity to clinch victory.

But Flushing Meadows has not seen the last of Serena just yet, with the 40-year-old back in action on Arthur Ashe on Friday for an eagerly anticipated third round singles clash with Australia's Ajla Tomljanovic.

(With input from agencies)