This paper describes a simple strategy to biochemically manipulate a surface at the nanoscale by enzyme dip-pen nanolithography using an endonuclease (DNase I) that is directly patterned on a self-assembled monolayer presenting a terminal oligonucleotide. Physisorbed nanopatterns of DNase I carried out nanoscale enzymology at the surface creating oligonucleotide patterns with the fidelity of the patterned enzyme because of the affinity of the enzyme for the immobilized, oligonucleotide substrate.