Still licking their wounds from Thursday night's Ryman Cup Final defeat in extra time against Charlton, the team led 2-1 going into the final 10 minutes at Brighton's American Express Elite Football Performance Centre before succumbing to two late goals.
Avilla Bergin and Bianca Baptiste went close for Spurs before Amy Taylor opened the scoring for Brighton on 26 minutes.
Wendy Martin struck low to level on 33 minutes and we were denied further by Faye Baker in the Brighton goal as it stayed 1-1 at the break.
Avilla fired us into the lead on the hour and the Seagulls - scrapping it out with Portsmouth and Charlton for the title - were forced to chase the game.
Toni-Anne Wayne kept out Sophie Perry and Lucy Somes struck the crossbar before goals from Emily Baker (82 minutes) and Kirsty Barton (88) turned it around for the home side.
It was Spurs' first defeat in the FAWPL in 2015 and they remain fifth in the table with a game in hand.
"The girls were a little deflated after the cup final, not because of the performance, but just what losing a final like that takes out of you," said manager, Karen Hills.
"It was a great response against one of the best teams in the league.
"We matched them all the way and could have been 3-1 up at half-time, but we didn't take our chances and they were more clinical than us.
"I couldn't fault the effort and application and I was so disappointed for the players because they've worked so hard in the last two games but have nothing to show for it.
"It just shows how far we've come to play so well in such high-profile matches.
"We now play Charlton again on Wednesday night and the players are all determined to get something out of the game."
Wednesday night's FAWPL match at third-placed Charlton is at Sporting Club Thamesmead, kick-off 7.45pm.
The ladies then have a clear weekend before facing Cardiff in the FAWPL on Sunday, May 3.
Spurs Ladies v Brighton: Toni-Anne Wayne; Rosie Kmita, Sophie McLean, Jenna Schillaci, Lucia Leon; Wendy Martin, Shannon Maloney, Riana Soobadoo (Leah Rawle, 80); Avilla Bergin (Eartha Pond, 80), Katie O'Leary, Bianca Baptiste. Unused sub: Leanne Mabey.
Congratulations to Spurs Ladies Reserves, who beat Petts Wood FC 3-1 in the Capital Intermediate Cup Final on Sunday.
SPURS LADIES MICROSITE