title: Human epidermal growth factor. Distinct roles of tyrosine 37 and arginine 41 in receptor binding as determined by site-directed mutagenesis and nuclear magnetic resonance spectroscopy.
abstract: Site-directed mutagenesis was employed to examine the function of two highly conserved residues, Tyr-37 and Arg-41, of human EGF (hEGF) in receptor binding. Both a conservative change to phenylalanine and a semi-conservative change to histidine at position 37 yield proteins with receptor affinity similar to wild-type hEGF. A non-conservative change to alanine results in a molecule with about 40% of the receptor affinity, indicating that an aromatic residue is not essential at this position. Both conservative (to lysine) and non-conservative (to alanine) substitutions at position 41 drastically reduced receptor binding to less than 0.5% of the wild-type activity. 1D-NMR data indicate that the replacement of Arg-41 by lysine does not significantly alter the native protein conformation. Thus, Arg-41 may be directly involved in ligand receptor interaction, whereas the side chain of Tyr-37, although possibly important structurally, is not essential for receptor binding.
authors.affiliation: Protein Engineering and Molecular Mutagenesis Program, Oak Ridge National Laboratory, Tennessee 37831-8077.
authors.forename: D A
authors.initials: DA
authors.lastname: Engler
authors.forename: G T
authors.initials: GT
authors.lastname: Montelione
authors.forename: S K
authors.initials: SK
authors.lastname: Niyogi
mesh_terms: D000595:Amino Acid Sequence; D001120:Arginine / Q000235:genetics / Q000378:metabolism*; D001667:Binding, Competitive; D004815:Epidermal Growth Factor / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009154:Mutation; D011487:Protein Conformation; D017459:Receptors, Amino Acid; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000235:genetics / Q000378:metabolism*
chemical_list: D017459:Receptors, Amino Acid; D011956:Receptors, Cell Surface; D014443:Tyrosine; D004815:Epidermal Growth Factor; D001120:Arginine