S1 pocket fingerprints of human and bacterial methionine aminopeptidases determined using fluorogenic libraries of substrates and phosphorus based inhibitors

► We demonstrated substrate specificity of human and E. coli methionine aminopeptidases. ► Screened MetAPs recognize methionine and other unnatural amino acids. ► We biochemically characterized the best substrates. ► We identified good phosphonate inhibitors for all tested methionine aminopeptidases. ► We provided information for further substrate and inhibitor design.