An early try from Leinster full back Rob Kearney and 20 points from the boot of Scarlets fly-half Stephen Jones should have been enough to see the Lions take the three-match series to a decider but Morne Steyn's 54-metre penalty secured series victory for the world champions with the final play of the match.
The Lions were superb throughout the first half as they started where they left off in Durban eight days earlier.
That first Test defeat had ended with the Lions crossing for two converted tries in the final 12 minutes and Ian McGeechan's men began Saturday's second encounter in similar fashion.
Jones put the first points on the board with a penalty after three minutes but not before Springbok flanker Schalk Burger had received a yellow card after appearing to make contact with the eye area of Leinster wing Luke Fitzgerald with just a minute played. While Sky Sports' commentators suggested that Burger had been lucky to escape with just a yellow, the Lions made the 50-cap flanker pay for his indiscretion by scoring a point a minute during his time in the sin bin.
If Jones' first penalty had given the Lions an early boost, Kearney's seventh-minute try lifted spirits even higher.
A magical offload from Jones gave Kearney possession 30 metres out, from where the he supplied an impressive finish. The presence of Ospreys wing Tommy Bowe kept JP Pietersen pegged to the left touchline, while Kearney's text-book decision to keep both hands firmly planted on the ball left opposite number Francois Steyn simply guessing as to what he would do next.
Kearney was in no doubt as to his own intentions, however, as he used Bowe's presence top open up the necessary gap and force his way straight to the tryline.
Jones then set the tone for an immaculate kicking display as he sent the difficult conversion straight through the uprights from just three metres in from the right-hand touchline.
The Lions continued to control the early stages, dominating territory and possession, but the Springboks soon showed a sign of things to come as they hit back in some style with 13 minutes on the clock.
Lineout ball just outside the Lions' 22 saw scrum-half du Preez drift across field with Pietersen slicing through the first line of defence on his way to his 11th try in South African colours.
Ruan Pienaar's conversion struck the far upright from just 10 metres to the left of posts but the Boks had at least reminded their own support exactly what they were capable of.
Jones added a second penalty on 16 minutes before trading three pointers with Francois Steyn prior to the end of the first 40 minutes - Jones through a cleverly-worked drop goal and Steyn through a long-range penalty attempt on the stroke of half-time.
The Lions entered the break with a 16-8 advantage but they were unable to hang on to that lead for the full 80 minutes.
Whereas the Lions had completely controlled the opening exchanges, the second period was a far more evenly-contested affair.
The boot of Jones kept the scoreboard ticking over for the Lions but ultimately two converted tries and two penalties in the final quarter would win the game for the Boks.
Former IRB World Player of the Year Bryan Habana claimed the first of those tries after 63 minutes, with replacement centre Jaque Fourie adding the second with five minutes remaining. Those scores alone would not have been enough had the home side continued with the erratic kicking form shown by both Ruan Pienaar and Francois Steyn but, unfortunately for the Lions, Morne Steyn proved to be a far more reliable points scorer.
The man who almost single-handed ensured the Bulls held off the challenge of the Crusaders in last month's Super 14 semi-final earned himself legendary status in the South African game despite this being just his second appearance in senior Springbok colours.
Steyn added the extras to Habana's try two minutes after his first penalty since replacing Pienaar, before again splitting the posts with a touchline conversion to Fourie's try that had given the Boks the lead for the very first time in the match.
A fifth successful penalty from five attempts from Jones appeared to have given the Lions hope of tying the series next week in Johannesburg as the score moved to 25 points apiece but Steyn had other ideas.
When Lions replacement Ronan O'Gara made contact with du Preez in the air after chasing an up and under, the whole of Loftus Versfeld held their breath. Referee Christophe Berdos awarded a penalty, the hooter sounded to signal the final play of the match, and Steyn held his nerve from inside his own half to bring an end to the Lions' hopes of a first series victory since 1997.
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