The interaction between sphalerite and pyrite was investigated by dissolution test, X-ray photoelectron spectroscopy (XPS), zeta potential measurement and density functional theory (DFT) calculation. Dissolution tests indicated that sphalerite dissolution was promoted due to the galvanic interaction between sphalerite and pyrite. The Zn²⁺ ion concentration increased with increasing pyrite content and dissolved time. XPS analysis results demonstrated that a new oxidation product was formed on the sphalerite surface in the presence of pyrite in a pulp. Zeta potential measurements showed that the isoelectric point of sphalerite increased from 3.3 to 5.4 due to galvanic interaction. DFT calculation results suggested that electron transfer from sphalerite to pyrite occurred when they contacted. The Zn 4s and S 3p states of sphalerite lost electrons. The Fe 4p and 4s of pyrite states obtained electrons, and Fe 3d and S 3s states lost a small number of electrons. The surface oxidation of sphalerite was promoted due to the interaction with pyrite, and the collectorless floatability of sphalerite decreased.