Distinct roles for ADAM10 and ADAM17 in ectodomain shedding of six EGFR ligands.
All ligands of the epidermal growth factor receptor (EGFR), which has important roles in development and disease, are released from the membrane by proteases. In several instances, ectodomain release is critical for activation of EGFR ligands, highlighting the importance of identifying EGFR ligand sheddases. Here, we uncovered the sheddases for six EGFR ligands using mouse embryonic cells lacking candidate-releasing enzymes (a disintegrin and metalloprotease [ADAM] 9, 10, 12, 15, 17, and 19). ADAM10 emerged as the main sheddase of EGF and betacellulin, and ADAM17 as the major convertase of epiregulin, transforming growth factor alpha, amphiregulin, and heparin-binding EGF-like growth factor in these cells. Analysis of adam9/12/15/17-/- knockout mice corroborated the essential role of adam17-/- in activating the EGFR in vivo. This comprehensive evaluation of EGFR ligand shedding in a defined experimental system demonstrates that ADAMs have critical roles in releasing all EGFR ligands tested here. Identification of EGFR ligand sheddases is a crucial step toward understanding the mechanism underlying ectodomain release, and has implications for designing novel inhibitors of EGFR-dependent tumors.
Sahin
Umut
U
Cell Biology Program, Sloan-Kettering Institute, Memorial Sloan-Kettering Cancer Center, Box 368, 1275 York Avenue, New York, NY 10021, USA.
Weskamp
Gisela
G
Kelly
Kristine
K
Zhou
Hong-Ming
HM
Higashiyama
Shigeki
S
Peschon
Jacques
J
Hartmann
Dieter
D
Saftig
Paul
P
Blobel
Carl P
CP
D051722:ADAM Proteins; D000072199:ADAM12 Protein; D000072198:ADAM17 Protein; D066258:Amphiregulin; D053829:Amyloid Precursor Protein Secretases; D000818:Animals; D016282:Aspartic Acid Endopeptidases; D066259:Betacellulin; D002478:Cells, Cultured; D019483:Disintegrins / Q000235:genetics / Q000378:metabolism; D066255:EGF Family of Proteins; D004622:Embryo, Mammalian / Q000033:anatomy & histology; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism*; D004815:Epidermal Growth Factor / Q000378:metabolism; D066260:Epiregulin; D066246:ErbB Receptors / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005838:Genotype; D006023:Glycoproteins / Q000378:metabolism; D066257:Heparin-binding EGF-like Growth Factor; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D008024:Ligands; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000378:metabolism; D011480:Protease Inhibitors / Q000378:metabolism; D017434:Protein Structure, Tertiary; D013755:Tetradecanoylphorbol Acetate / Q000378:metabolism; D013876:Thiophenes / Q000378:metabolism; D016211:Transforming Growth Factor alpha / Q000378:metabolism
D066258:Amphiregulin; C587849:Areg protein, mouse; D066259:Betacellulin; C587852:Btc protein, mouse; D019483:Disintegrins; D066255:EGF Family of Proteins; D066260:Epiregulin; C587881:Ereg protein, mouse; D006023:Glycoproteins; C587884:Hbegf protein, mouse; D066257:Heparin-binding EGF-like Growth Factor; D036341:Intercellular Signaling Peptides and Proteins; D008024:Ligands; D008565:Membrane Proteins; D009124:Muscle Proteins; D011480:Protease Inhibitors; D013876:Thiophenes; D016211:Transforming Growth Factor alpha; D010649:Phenylalanine; D004815:Epidermal Growth Factor; C080985:batimastat; D066246:ErbB Receptors; D053829:Amyloid Precursor Protein Secretases; D010450:Endopeptidases; D016282:Aspartic Acid Endopeptidases; C509282:Bace1 protein, mouse; D051722:ADAM Proteins; D000072199:ADAM12 Protein; C096115:Adam12 protein, mouse; C490902:Adam15 protein, mouse; C498316:Adam9 protein, mouse; D008666:Metalloendopeptidases; D000072198:ADAM17 Protein; C000606463:Adam17 protein, mouse; D013755:Tetradecanoylphorbol Acetate