The effects of two C(60) derivatives: C(60)[C(COOH)(2)](3) and C(60)(NH(2))(x)(OH)(y), have been investigated on the bioluminescence of humanized Gaussia luciferase (hGluc) for the first time. Utilizing the dual properties of carboxyl C(60) with luminescence quenching and free radicals scavenging, a novel BRET system was constructed for protease detection with high sensitivity.