Zdenek Ondrasek came off the bench on his international debut to earn the Czech Republic a 2-1 win at home to England, denying the Three Lions qualification for Euro 2020.
Harry Kane's fifth-minute penalty - the England captain's 27th international goal - put Gareth Southgate's side in front early on Friday, yet the hosts hit back within five minutes, reports Goal.
Centre-back Jakub Brabec profited on slack set-piece marking to level the scores and, as the game appeared to be drifting towards a draw, the Czech Republic struck again.
With five minutes to go, Lukas Masopust raided down the right and pulled a low cross back for striker Ondrasek, who confidently beat Jordan Pickford to snatch all three points.
Kane missed from the spot against Kosovo last month but he converted a fifth-minute penalty in Prague.
Raheem Sterling's darting run down the left was ended by Masopust's clumsy trip and Kane fired the resulting spot-kick down the middle, though the hosts soon drew level.
A corner found Brabec at the back post with Declan Rice having slipped and the defender bundled home his first international goal, beating Danny Rose on the line.
Goalkeeper Tomas Vaclik was quickly off his line to prevent Sterling rounding him in the 57th minute, then Jordan Pickford was called into action to acrobatically deny Masopust.
Alex Kral forced another good save from Pickford in the closing stages but England did not heed the warning, Ondrasek slamming home with his first shot in international football.