Detection of post-translational modifications in single peptides using electron tunnelling currents

Post-translational modifications alter the properties of proteins through the cleavage of peptide bonds or the addition of a modifying group to one or more amino acids(1-7). These modifications allow proteins to perform their primary biological functions(8-11), but single-protein studies of post-translational modifications have been hindered by a lack of suitable analysis methods(12-16). Here, we show that single amino acids can be identified using electron tunnelling currents measured as the individual molecules pass through a nanoscale gap between electrodes. We identify 12 different amino acids and the post-translational modification phosphotyrosine, which is involved in the process that switches enzymes on and off(17-20). Furthermore, we show that the conductance measurements can be used to partially sequence peptides of an epidermal growth factor receptor substrate, and can discriminate a peptide from its phosphorylated variant.