Insights into the bonding of As(V) at the metal oxide/aqueous interface can further our understanding of its fate and transport in the environment. The motivation of this work is to explore the interfacial configuration of As(V) on single crystal rutile (110) using grazing-incidence X-ray absorption fine structure spectroscopy (GI-XAFS) and planewave density functional calculations with on-site repulsion (DFT+U). In contrast to the commonly considered corner-sharing bidentate binuclear structure, tetrahedral As(V) binds as an edge/corner-sharing tridentate binuclear complex on rutile (110), as evidenced by observation of three As-Ti distances at 2.83, 3.36, and 4.05 angstrom. In agreement with the GI-XAFS analysis, our DFT+U calculations for this configuration resulted in the lowest adsorption energy among five possible alternatives. In addition, the electron density difference further demonstrated the transfer of charge between surface Ti atoms and O atoms in AsO4. This charge transfer consequently induced the formation of a chemical bond, which is also confirmed by the partial density of states analysis. Our results may shed new light on coupling the GI-XAFS and DFT approaches to explore molecular-scale adsorption mechanisms on single crystal surfaces.