Structure-function analysis of epidermal growth factor: site directed mutagenesis and nuclear magnetic resonance.
The role of leucine-47 in determining the structure and activity of human epidermal growth factor was examined using site-directed mutagenesis. Wild type protein and four variants in which Leu47 was replaced by valine, glutamate, aspartate and alanine were produced from yeast. 1H NMR experiments demonstrated that substitution of Leu47 had little effect on the protein structure. The observed reduction in receptor binding affinity caused by the substitutions could thus be attributed to perturbation of a residue directly involved in receptor interactions.
Dudgeon
T J
TJ
Department of Biochemistry, University of Oxford, UK.
Cooke
R M
RM
Baron
M
M
Campbell
I D
ID
Edwards
R M
RM
Fallon
A
A
D001667:Binding, Competitive; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003001:Cloning, Molecular; D004815:Epidermal Growth Factor / Q000235:genetics / Q000378:metabolism*; D066246:ErbB Receptors / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D007930:Leucine; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D009154:Mutation; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013329:Structure-Activity Relationship
D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D007930:Leucine