Ligand-induced functions of the epidermal growth factor receptor require the positively charged region asymmetrically distributed across plasma membrane.
Many plasma membrane proteins, including the epidermal growth factor (EGF) receptor, possess basic regions on the cytoplasmic surface of the membrane. To examine the function of these positively charged regions, we constructed mutated EGF receptor genes lacking this region by substitution of the basic amino acid residues with 3 approximately 8 neutral Asn residues, or by their complete deletion. There was no significant difference in the affinities for EGF of the wild-type and mutant receptors which are produced in rodent fibroblasts through transfection. However, EGF-induced tyrosine phosphorylation of the receptor was strongly inhibited by removal of the 3 approximately 8 positively charged residues. On addition of EGF, cells expressing the mutant EGF receptors did not show morphological changes, whereas cells expressing the wild-type receptor did. These findings suggest that the positively charged regions of membrane proteins that are asymmetrically distributed on the cytoplasmic surface of the membrane may be required for the functions of membrane proteins in general.
Yamane
K
K
Biology Division, National Cancer Center Research Institute, Tokyo, Japan.
Toyoshima
C
C
Nishimura
S
S
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002872:Chromosome Deletion; D004815:Epidermal Growth Factor / Q000378:metabolism / Q000494:pharmacology*; D066246:ErbB Receptors / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D008024:Ligands; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides; D010766:Phosphorylation; D010957:Plasmids; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D015183:Restriction Mapping; D014162:Transfection
D008024:Ligands; D009838:Oligodeoxyribonucleotides; D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D011505:Protein-Tyrosine Kinases