The study of biomolecular motors represents a rapidly and progressing field of nanobiotechnology. Here, we present a simple method for patterning myosin II on a microstructured surface. Our findings indicate that UV/ozone treatment can be used to alter the hydrophobicity of trimethyl-chloro-silane (TMCS) coated glass surfaces, to alter protein binding, and effectively produce localized motor activity. Taken together, these data suggest that photoreactive patterning may be useful for the selective localization of functional myosin II motor tracks.