Flyhalf Michael Hobbs and All Black loose forward Adam Thomson have not been considered for selection for the Super 14 match against the Bulls in Pretoria on Saturday.
The duo have been axed, after they failed to comply with the team curfew following the Stormers game last week.
The Highlanders had a curfew following their match in Cape Town on the weekend, but Highlanders head coach Glenn Moore said the players didn't get back to the team hotel on time.
They weren't involved in any other incidents.
Moore said that while suspending the players was a critical decision that impacted on the whole team, it was in the end a straightforward decision to make.
"Each week you have to earn the right to play for this team. This slip up, albeit out of character for these two guys, means they don't get the opportunity to play this week," Moore said.
Moore said Hobbs and Thomson would stay with the team in Pretoria to help them prepare for the match against the Bulls.
Steven Setephano starts at No.8 in place of Thomson, while Mat Berquist will start at No.10.
The team is also without injured captain Jimmy Cowan, with Sean Romans starting at No.9 and Jamie Mackintosh taking over the captaincy. Cowan's brother Scott comes onto the bench.
The Highlanders return to New Zealand on Sunday following the match.
This is their final game of the South African leg of the competition.
Highlanders: 15 Israel Dagg, 14 Ben Smith, 13 Kenny Lynn, 12 Jason Shoemark, 11 Fetu'u Vainikolo, 10 Mat Berquist, 9 Sean Romans, 8 Steven Setephano, 7 Alando Soakai, 6 Tim Boys, 5 Tom Donnelly, 4 Josh Bekhuis, 3 Chris King, 2 Jason Macdonald, 1 Jamie Mackintosh (captain).
Replacements: 16 Jason Rutledge, 17 Bronson Murray, 18 Hayden Triggs, 19 John Hardie, 20 Scott Cowan, 21 Robbie Robinson, 22 James Paterson.
Date: Saturday, March 13
Venue: Loftus Versfeld, Pretoria
Kick-off: 17.05 (15.05 GMT)
Referee: Jonathan Kaplan (South Africa)
Assistant referees: Pro Legoete (South Africa), Christie du Preez (South Africa)
TMO: Johann Meuwesen (South Africa)
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