This work proposes a quenched photoelectrochemical sensing method for highly selective and sensitive detection of protein via Energy Transfer (ET) effect between the AuNPs and CdS:Mn quantum dots. This detection was performed on a sandwich-type aptamer sensing interface. Chitosan modified CdS:Mn/TiO2/ITO electrode was used to immobilize capture DNA (S1) via -CONH- bond. In the presence of target protein, AuNPs labeled DNA (AuNPs-S2) was further bonded to the protein to fabricate sandwich sensing platform, which forced the AuNPs away from the electrode surface. In this state, the photocurrent was greatly depressed, mainly due to two factors: (a) the ET effect produced by interparticle distance between CdS:Mn and AuNPs; (b) the steric hindrance of AuNPs-S2 partly obstructs the diffusion of the electron donor. The photocurrent decreased with the increasing concentration of the target protein. Using thrombin as a target, this sensitized method showed a detectable range of 0.1 pM to 8 nM and a detection limit of 30 fM. It possessed high selectivity and good stability for detection of thrombin. This method is extremely flexible and can be extended to varieties of protein targets.