Andy Murray saw one of his bids for Wimbledon silverware prove fruitless as the Briton and France's Pierre-Hugues Herbert lost in the men's doubles.
Murray and Herbert were beaten 6-7 (4-7) 6-4 6-2 6-3 by Croatian sixth seeds Nikola Mektic and Franko Skugor.
All is not lost for the Scot, however, as he has another chance alongside American great Serena Williams in the mixed doubles.
The illustrious pair face Andreas Mies and Alexa Guarachi later on Saturday.
Murray and Williams, two of the sport's most recognisable players, will meet their German-Chilean opponents on a court to be decided after 17:30 BST.
Regardless of what happens in that match, Murray's overriding emotion of returning to Wimbledon - six months after he had a hip resurfacing operation - will be happiness at being competitive.
The 32-year-old Scot made a triumphant return when he and Herbert earned a comeback win over Romania's Marius Copil and France's Ugo Humbert, recovering from a slow start to enthral a boisterous Court One on Thursday by winning in four sets.
But this time the mood on a packed court two, one of the smaller show courts at the All England Club, faltered as Murray and Herbert's second-round match swung in the opposite direction.
The partnership failed to ignite in the same way that Murray's triumphant pairing with Spain's Feliciano Lopez did at Queens, with an almost innate understanding between doubles specialists Mektic and Skugor proving too much.
Initially it looked like it could be another positive outcome for Murray and Herbert, who edged an even first set after a crisp cross-court backhand winner from Murray swung the tie-break in their favour.
But 28-year-old Herbert, who has won all four Grand Slam doubles titles after success with his previous partner Nicolas Mahut, continued to struggle with his returning game in the second set and then crucially saw his serve taken for their opponents to level.
From that point Mektic and Skugor took control as Murray and Herbert's service game waned, the Croatians breaking three more times in the next two sets to reach the third round.
'Doubt we'll see Murray & Herbert in tandem again' - analysis
Peter Fleming, seven-time Grand Slam champion on BBC TV
Mektic and Skugor were good value for their win, after a sluggish start they did play some superb tennis.
We probably won't see a reprise of the Murray-Herbert tandem anytime soon. They just didn't gel.
Andrew Castle, former British number one on BBC TV
I think Herbert will be disappointed with himself more than Murray, who has done more than enough in this grass court season and will be delighted to be back.
Herbert went a little flat in that second set if truth be told.