title: Cell surface annexins regulate ADAM-mediated ectodomain shedding of proamphiregulin.
abstract: A disintegrin and metalloproteinase (ADAM) is a family of enzymes involved in ectodomain shedding of various membrane proteins. However, the molecular mechanism underlying substrate recognition by ADAMs remains unknown. In this study, we successfully captured and analyzed cell surface transient assemblies between the transmembrane amphiregulin precursor (proAREG) and ADAM17 during an early shedding phase, which enabled the identification of cell surface annexins as components of their shedding complex. Annexin family members annexin A2 (ANXA2), A8, and A9 interacted with proAREG and ADAM17 on the cell surface. Shedding of proAREG was increased when ANXA2 was knocked down but decreased with ANXA8 and A9 knockdown, because of enhanced and impaired association with ADAM17, respectively. Knockdown of ANXA2 and A8 in primary keratinocytes altered wound-induced cell migration and ultraviolet B-induced phosphorylation of epidermal growth factor receptor (EGFR), suggesting that annexins play an essential role in the ADAM-mediated ectodomain shedding of EGFR ligands. On the basis of these data, we propose that annexins on the cell surface function as "shedding platform" proteins to determine the substrate selectivity of ADAM17, with possible therapeutic potential in ADAM-related diseases.
authors.affiliation: Department of Biochemistry and Molecular Genetics, Ehime University Graduate School of Medicine, Shitsukawa, Toon, Ehime, Japan.
authors.forename: Hironao
authors.initials: H
authors.lastname: Nakayama
authors.forename: Shinji
authors.initials: S
authors.lastname: Fukuda
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Inoue
authors.forename: Hisayo
authors.initials: H
authors.lastname: Nishida-Fukuda
authors.forename: Yuji
authors.initials: Y
authors.lastname: Shirakata
authors.forename: Koji
authors.initials: K
authors.lastname: Hashimoto
authors.forename: Shigeki
authors.initials: S
authors.lastname: Higashiyama
mesh_terms: D051722:ADAM Proteins / Q000378:metabolism*; D000072198:ADAM17 Protein; D000595:Amino Acid Sequence; D066258:Amphiregulin; D017302:Annexins / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement; D017077:Culture Media, Conditioned; D066255:EGF Family of Proteins; D066246:ErbB Receptors / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry / Q000378:metabolism*; D015603:Keratinocytes / Q000378:metabolism / Q000502:physiology / Q000528:radiation effects; D010766:Phosphorylation; D061251:Primary Cell Culture; D011485:Protein Binding; D011498:Protein Precursors / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D059748:Proteolysis; D034622:RNA Interference; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D059008:Time-Lapse Imaging; D015533:Transcriptional Activation; D014466:Ultraviolet Rays
chemical_list: C587848:AREG protein, human; D066258:Amphiregulin; D017302:Annexins; D017077:Culture Media, Conditioned; D066255:EGF Family of Proteins; D006023:Glycoproteins; D036341:Intercellular Signaling Peptides and Proteins; D011498:Protein Precursors; D066246:ErbB Receptors; D051722:ADAM Proteins; D000072198:ADAM17 Protein; C000606462:ADAM17 protein, human; D013755:Tetradecanoylphorbol Acetate