ADAM17 is regulated by a rapid and reversible mechanism that controls access to its catalytic site

Sylvain M Le Gall; Thorsten Maretzky; Priya D. A Issuree; Xiao-Da Niu; Karina Reiss; Paul Saftig; Rama Khokha; Daniel Lundell; Carl P Blobel

doi:10.1242/jcs.069997

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ADAM17 is regulated by a rapid and reversible mechanism that controls access to its catalytic site

Journal article

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ADAM17 is regulated by a rapid and reversible mechanism that controls access to its catalytic site

Sylvain M Le Gall, Thorsten Maretzky, Priya D. A Issuree, Xiao-Da Niu, Karina Reiss, Paul Saftig, Rama Khokha, Daniel Lundell and Carl P Blobel

Journal of cell science, Vol.123(22), pp.3913-3922

11/15/2010

DOI: 10.1242/jcs.069997

PMCID: PMC2972273

PMID: 20980382

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Abstract

Protein ectodomain shedding is crucial for cell–cell interactions because it controls the bioavailability of soluble tumor necrosis factor-α (TNFα) and ligands of the epidermal growth factor (EGF) receptor, and the release of many other membrane proteins. Various stimuli can rapidly trigger ectodomain shedding, yet much remains to be learned about the identity of the enzymes that respond to these stimuli and the mechanisms underlying their activation. Here, we demonstrate that the membrane-anchored metalloproteinase ADAM17, but not ADAM10, is the sheddase that rapidly responds to the physiological signaling pathways stimulated by thrombin, EGF, lysophosphatidic acid and TNFα. Stimulation of ADAM17 is swift and quickly reversible, and does not depend on removal of its inhibitory pro-domain by pro-protein convertases, or on dissociation of an endogenous inhibitor, TIMP3. Moreover, activation of ADAM17 by physiological stimuli requires its transmembrane domain, but not its cytoplasmic domain, arguing against inside–out signaling via cytoplasmic phosphorylation as the underlying mechanism. Finally, experiments with the tight binding hydroxamate inhibitor DPC333, used here to probe the accessibility of the active site of ADAM17, demonstrate that this inhibitor can quickly bind to ADAM17 in stimulated, but not quiescent cells. These findings support the concept that activation of ADAM17 involves a rapid and reversible exposure of its catalytic site.

Cell–cell interactions

ADAM17

EGF-receptor

Details

Title: Subtitle: ADAM17 is regulated by a rapid and reversible mechanism that controls access to its catalytic site
Creators: Sylvain M Le Gall - Arthritis and Tissue Degeneration Program, Hospital for Special Surgery
Thorsten Maretzky - Arthritis and Tissue Degeneration Program, Hospital for Special Surgery
Priya D. A Issuree - Arthritis and Tissue Degeneration Program, Hospital for Special Surgery
Xiao-Da Niu - Department of Respiratory Care and Immunology, Merck Research Laboratories
Karina Reiss - Clinical Research Unit, Department of Dermatology, Christian-Albrechts-University
Paul Saftig - Biochemical Institute, Christian-Albrechts-University
Rama Khokha - Ontario Cancer Institute, University of Toronto
Daniel Lundell - Department of Respiratory Care and Immunology, Merck Research Laboratories
Carl P Blobel - Arthritis and Tissue Degeneration Program, Hospital for Special Surgery
Resource Type: Journal article
Publication Details: Journal of cell science, Vol.123(22), pp.3913-3922
DOI: 10.1242/jcs.069997
PMID: 20980382
PMCID: PMC2972273
NLM abbreviation: J Cell Sci
ISSN: 0021-9533
eISSN: 1477-9137
Publisher: Company of Biologists
Language: English
Date published: 11/15/2010
Academic Unit: Infectious Diseases; Internal Medicine
Record Identifier: 9984094376102771

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