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Photocatalytic methane conversion under ambient conditions holds immense promise for sustainable use of methane resources, but incomplete knowledge of reaction mechanisms hampers the development of engineering strategies for methane photocatalysis. Here, combining real-time mass spectrometry and operando infrared absorption spectroscopy with ab initio molecular dynamics simulations, the authors explore the role of interfacial water in C–H activation during the photocatalytic conversion of methane.