Recognition of an antiparallel beta-sheet structure of human epidermal growth factor by its receptor. Site-directed mutagenesis studies of Ala-30 and Asn-32.
The Ala-30 and Asn-32 residues involved in the major antiparallel beta-sheet structure of human epidermal growth factor (hEGF) were substituted with various amino acid residues, and the receptor-binding affinities of the nine variant hEGFs were determined by the use of human KB cells. The Ala-30----Arg, Ala-30----His and Ala-30----Phe substitutions drastically reduced the binding affinity, suggesting that the side chain in position 30 of Ala-30 of hEGF is required to be small for the receptor binding. The Asn-32----Asp substitution significantly reduced the binding affinity, while the Asn-32----His variant could bind to the receptor as well as to the wild-type hEGF. Therefore, it seems to be important for receptor binding that the side chain in position 32 does not have a negative charge but does have an NH group. Thus, we propose that, in the ligand-receptor complex, the receptor recognizes, on one side of the antiparallel beta-sheet structure of hEGF, a wider contact area than previously suggested.
Koide
H
H
Department of Biophysics and Biochemistry, Faculty of Science, University of Tokyo, Japan.
Muto
Y
Y
Kasai
H
H
Hoshi
K
K
Takusari
H
H
Kohri
K
K
Takahashi
S
S
Sasaki
T
T
Tsukumo
K
K
Miyake
T
T
D000409:Alanine / Q000378:metabolism; D001216:Asparagine / Q000378:metabolism; D001667:Binding, Competitive; D004815:Epidermal Growth Factor / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D066246:ErbB Receptors / Q000378:metabolism*; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation
D004815:Epidermal Growth Factor; D001216:Asparagine; D066246:ErbB Receptors; D000409:Alanine