Mexico skipper Rafael Marquez scored an 89th-minute winner as the United States crashed to their first World Cup qualifying defeat on home soil in 15 years with a 2-1 loss.
Marquez, 37, headed home a Miguel Layun corner in the dying minutes of a pulsating battle to end the USA's long record of dominance at Columbus's Mapfre Stadium, where they had beaten Mexico in the past four World Cup cycles.
Marquez's goal came after the US had clawed their way back into the contest with a Bobby Wood equaliser on 49 minutes after Layun's 20th-minute goal put Mexico 1-0 up.
Jurgen Klinsmann's side had looked the likeliest to take all three points but instead it was Marquez who struck to snatch Mexico's first ever qualifying win over the United States at their Columbus stronghold.
"There's nothing I can complain about the team, they gave everything they had, they kept the tempo high, we had enough chances to put it away, but we didn't," US coach Klinsmann said afterwards.
Mexico goalscorer Layun said 'El Tri' had been worthy winners.
"We deserved the three points. It puts us in a good position," Layun said. "We were focused on this. That's why we got the three points."
The result gives Mexico the early initiative in the CONCACAF region's six-team final round of qualifying for the 2018 World Cup in Russia.
The United States now face a testing trip to Costa Rica on Tuesday needing to pick themselves up after a shattering loss.
The match took place against the backdrop of the US presidential election won by Donald Trump on Tuesday after a campaign which included rhetoric against Mexican immigrants.
However fans from both sides mingled before kick-off, adamant that politics would not poison the atmosphere.
Mexico had the better of a frenetic first half, rattling US nerves after only 10 minutes when Jesus Corona cut in from the left and saw his shot tipped against the woodwork after a diving save from Tim Howard.
At the other end, the US threatened sporadically but were unable to find the breakthrough, with Jozy Altidore's 20th minute header easily gathered by Mexico goalkeeper Alfredo Talavera.
Moments later Mexico took the lead. Again it was an attack that began down the left flank, Layun pouncing on a loose ball near the edge of the area and driving a low shot which took a slight deflection as it flew past Howard.
The US were rocked and Mexico almost doubled their tally five minutes later when Corona's cross was met with a glancing header from Carlos Vela which crashed back off the crossbar.
The best US chance of the half came in the 30th minute when a quick free-kick from captain Michael Bradley sowed confusion and the ball fell to Christian Pulisic only for the Borussia Dortmund's teenager's first touch to let him down.
The US were forced into an injury substitution on 39 minutes when Howard was replaced by Brad Guzan after appearing to pull his groin during a goal kick.
The US upped the tempo at the start of the second half and their more energetic play paid off immediately with an equaliser.
Altidore turned away Hugo Ayala brilliantly and powered upfield before releasing Wood, who stabbed a low deflected shot past Talavera.
The US pressed forward relentlessly looking for a winner, Mexico won a late corner and Marquez glanced home to deliver a killer sucker punch.