Identification of a 13 Amino Acid Peptide Mimetic of Erythropoietin and Description of Amino Acids Critical for the Mimetic Activity of EMP1

To obtain information about the functional importance of amino acids required for effective erythropoietin (EPO) mimetic action, the conserved residues of a peptide mimetic of EPO, recently discovered by phage display, were subjected to an alanine replacement strategy. Further, to identify a minimal mimetic peptide sequence, a series of truncation peptides has been generated. One EPO mimetic peptide sequence, EMP1, was targeted and more than 25 derivatives of this sequence were evaluated for their ability to compete with [125I]EPO for receptor binding and for their ability to support the proliferation of two EPO-responsive cell lines. Two hydrophobic amino acids, Tyr4 and Trp13, appear essential for mimetic action, and aromatic residues appear to be important at these sites. These findings are consistent with the previously reported X-ray crystal structure of EMP1 complexed with the extracellular domain of the EPO receptor (EPO binding protein; EBP). In our efforts to define the structural elements requir...