Tottenham's Champions League ambitions came crashing down as Juventus produced two goals inside three second-half minutes to decide the destiny of their last-16 tie.
Son Heung-min's scrambled finish six minutes before the interval strengthened Spurs' position at Wembley after they recovered from two goals down to draw the first leg in Turin 2-2.
In an eventful first 45 minutes, Son and Harry Kane had wasted chances for the hosts, while Juventus were denied a clear penalty when Douglas Costa was brought down by Jan Vertonghen.
Tottenham appeared to be in control until Gonzalo Higuain turned in Sami Khedira's flick to give Juventus a 64th-minute lifeline - which they capitalised on ruthlessly and Paulo Dybala raced clear for the decisive goal three minutes later.
Mauricio Pochettino's side fought to revive their chances but to no avail, coming agonisingly close in the dying moments when Kane's header hit the inside of the post but bounced right on the goal-line.
For Spurs, who last reached the Champions League quarter-finals in 2010-11, it was a first defeat in 18 matches, while Juventus are unbeaten since 22 November - a run of 20 games.