An unsymmetrical di-copper complex, ([Cu2(TPMAN)(μ-OH)(H2O)]3+, was prepared and used for electrocatalytic water oxidation in neutral conditions. This complex is a stable and efficient homogeneous catalyst during the electrocatalytic oxygen evolution process ( kcat = 0.78 s-1) with 780 mV onset overpotential in 0.1 M phosphate buffer (pH 7.0). The water oxidation mechanism of the unsymmetrical catalyst [Cu2(TPMAN)(μ-OH)]3+ exhibits different behaviors than that of [Cu2(BPMAN)(μ-OH)]3+, such as two redox steps with different pH dependences, a significant kinetic isotope effect, and buffer concentration dependence. All these changes were ascribed to the open site on the Cu center that is formed by removal of the hemilabile pyridyl site, which acts as an intramolecular proton acceptor to assist the O-O bond formation step.