The Cysteine-Rich Domain Regulates ADAM Protease Function in vivo

• Published in: The Journal of cell biology Vol. 159; no. 5; pp. 893 - 902
• Main Authors: Smith, Katherine M., Gaultier, Alban, Cousin, Helene, Alfandari, Dominique, White, Judith M., DeSimone, Douglas W.
• Format: Journal Article
• Language: English
• Published: United States Rockefeller University Press 09-12-2002; The Rockefeller University Press
• Subjects: ADAM Proteins; Amino Acid Sequence; Animal glands; Animals; Binding Sites; Cell adhesion; Cell Membrane - chemistry; Cells; Cellular biology; Cements; Chimera - genetics; Chimeras; Cysteine - chemistry; Disintegrins - chemistry; Disintegrins - genetics; Disintegrins - metabolism; Embryo, Nonmammalian - metabolism; Embryos; Exocrine Glands - embryology; Exocrine Glands - pathology; Extracellular Matrix - metabolism; Fertilins; Gene Expression Regulation, Developmental; Gene Expression Regulation, Enzymologic; Hyperplasia; Mathematical functions; Membrane Glycoproteins - chemistry; Membrane Glycoproteins - metabolism; Membrane Proteins - chemistry; Membrane Proteins - genetics; Membrane Proteins - metabolism; Membrane Proteins - physiology; Membranes; Messenger RNA; Metalloendopeptidases - chemistry; Metalloendopeptidases - genetics; Metalloendopeptidases - metabolism; Metalloendopeptidases - physiology; Models, Biological; Molecular Sequence Data; Mutagenesis, Site-Directed; Neural Crest - cytology; Neural Crest - metabolism; Protein Structure, Tertiary; Proteins; RNA; RNA, Messenger - metabolism; Sequence Alignment; Xenopus laevis - embryology; Xenopus laevis - genetics; Xenopus laevis - metabolism; Xenopus Proteins
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.200206023

ADAMs are membrane-anchored proteases that regulate cell behavior by proteolytically modifying the cell surface and ECM. Like other membrane-anchored proteases, ADAMs contain candidate "adhesive" domains downstream of their metalloprotease domains. The mechanism by which membrane-anchored cell surface proteases utilize these putative adhesive domains to regulate protease function in vivo is not well understood. We address this important question by analyzing the relative contributions of downstream extracellular domains (disintegrin, cysteine rich, and EGF-like repeat) of the ADAM13 metalloprotease during Xenopus laevis development. When expressed in embryos, ADAM13 induces hyperplasia of the cement gland, whereas ADAM10 does not. Using chimeric constructs, we find that the metalloprotease domain of ADAM10 can substitute for that of ADAM13, but that specificity for cement gland expansion requires a downstream extracellular domain of ADAM13. Analysis of finer resolution chimeras indicates an essential role for the cysteine-rich domain and a supporting role for the disintegrin domain. These and other results reveal that the cysteine-rich domain of ADAM13 cooperates intramolecularly with the ADAM13 metalloprotease domain to regulate its function in vivo. Our findings thus provide the first evidence that a downstream extracellular adhesive domain plays an active role in regulating ADAM protease function in vivo. These findings are likely relevant to other membrane-anchored cell surface proteases.