Accelerated degradation of 160 kDa epidermal growth factor (EGF) receptor precursor by the tyrosine kinase inhibitor herbimycin A in the endoplasmic reticulum of A431 human epidermoid carcinoma cells.
The effect of herbimycin A on the biosynthesis of epidermal growth factor (EGF) receptor was examined in human epidermoid carcinoma A431 cells. Cells were pulse-labelled with [35S]methionine, and EGF receptor biosynthesis was quantified by immunoprecipitation using a monoclonal anti-(EGF receptor) antibody. In the presence of herbimycin A, an immature 160 kDa EGF receptor precursor accumulated in 1 h and disappeared completely in 4 h. Pulse-labelled 160 kDa receptor precursor in the absence of herbimycin A, however, was converted normally into a 170 kDa one by chase with herbimycin A. Herbimycin A affected neither the synthesis of the secreted form of EGF receptor devoid of cytoplasmic domain, nor that of the transferrin receptor in A431 cells. The herbimycin A-induced degradation of 160 kDa EGF receptor precursor was not inhibited by an inhibitor of lysosomal enzymes, NH4Cl. Endoglycosidase H digestion of the 160 kDa precursor converted it into the deglycosylated 130 kDa precursor peptide. These results suggested that herbimycin A selectively acted on the EGF receptor precursor during the synthesis of the 160 kDa form, probably on the cytoplasmic domain, to form an aberrant molecule which was subjected to rapid degradation in the endoplasmic reticulum.
Department of Bioactive Molecules, National Institute of Health, Tokyo, Japan.
Y
Y
Murakami
S
S
Mizuno
Y
Y
Uehara
D016227:Benzoquinones; D002294:Carcinoma, Squamous Cell / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism; D066246:ErbB Receptors / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D047029:Lactams, Macrocyclic; D008970:Molecular Weight; D011498:Protein Precursors / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D011809:Quinones / Q000494:pharmacology*; D011990:Receptors, Transferrin / Q000096:biosynthesis / Q000187:drug effects; D017828:Rifabutin / Q000031:analogs & derivatives; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism
D016227:Benzoquinones; D047029:Lactams, Macrocyclic; D011498:Protein Precursors; D011809:Quinones; D011990:Receptors, Transferrin; D017828:Rifabutin; C020754:herbimycin; D066246:ErbB Receptors; D011505:Protein-Tyrosine Kinases