The distance between electrodes in a tunnel junction cannot be determined from the external movement applied to the electrodes because of interfacial forces that distort the electrode geometry at the nanoscale. These distortions become particularly complex when molecules are present in the junction, as demonstrated here by measurements of the AC response of a molecular junction over a range of conductivities from microsiemens to picosiemens. Specific chemical interactions within the junction lead to distinct features in break-junction data, and these have been used to determine the electrode separation in a junction functionalized with 4(5)-(2-mercaptoethyl)-1H-imidazole-2-carboxamide, a reagent developed for reading DNA sequences.