The Cytoplasmic Domain of A Disintegrin and Metalloproteinase 10 (ADAM10) Regulates Its Constitutive Activity but Is Dispensable for Stimulated ADAM10-dependent Shedding

Thorsten Maretzky; Astrid Evers; Sylvain Le Gall; Rolake O Alabi; Nancy Speck; Karina Reiss; Carl P Blobel

doi:10.1074/jbc.M114.603753

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The Cytoplasmic Domain of A Disintegrin and Metalloproteinase 10 (ADAM10) Regulates Its Constitutive Activity but Is Dispensable for Stimulated ADAM10-dependent Shedding

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The Cytoplasmic Domain of A Disintegrin and Metalloproteinase 10 (ADAM10) Regulates Its Constitutive Activity but Is Dispensable for Stimulated ADAM10-dependent Shedding

Thorsten Maretzky, Astrid Evers, Sylvain Le Gall, Rolake O Alabi, Nancy Speck, Karina Reiss and Carl P Blobel

The Journal of biological chemistry, Vol.290(12), pp.7416-7425

03/20/2015

DOI: 10.1074/jbc.M114.603753

PMCID: PMC4367251

PMID: 25605720

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Abstract

The membrane-anchored metalloproteinase a disintegrin and metalloprotease 10 (ADAM10) is required for shedding of membrane proteins such as EGF, betacellulin, the amyloid precursor protein, and CD23 from cells. ADAM10 is constitutively active and can be rapidly and post-translationally enhanced by several stimuli, yet little is known about the underlying mechanism. Here, we use ADAM10-deficient cells transfected with wild type or mutant ADAM10 to address the role of its cytoplasmic and transmembrane domain in regulating ADAM10-dependent protein ectodomain shedding. We report that the cytoplasmic domain of ADAM10 negatively regulates its constitutive activity through an ER retention motif but is dispensable for its stimulated activity. However, chimeras with the extracellular domain of ADAM10 and the transmembrane domain of ADAM17 with or without the cytoplasmic domain of ADAM17 show reduced stimulated shedding of the ADAM10 substrate betacellulin, whereas the ionomycin-stimulated shedding of the ADAM17 substrates CD62-L and TGFα is not affected. Moreover, we show that influx of extracellular calcium activates ADAM10 but is not essential for its activation by APMA and BzATP. Finally, the rapid stimulation of ADAM10 is not significantly affected by incubation with proprotein convertase inhibitors for up to 8 h, arguing against a major role of increased prodomain removal in the rapid stimulation of ADAM10. Thus, the cytoplasmic domain of ADAM10 negatively influences constitutive shedding through an ER retention motif, whereas the cytoplasmic domain and prodomain processing are not required for the rapid activation of ADAM10-dependent shedding events. Background: Release of membrane proteins from the cell surface by the metalloproteinase ADAM10 is post-translationally regulated. Results: The cytoplasmic domain of ADAM10 negatively controls its constitutive activity but is dispensable for its stimulation. Conclusion: Post-translational stimulation of ADAM10 requires its transmembrane and/or extracellular domain. Significance: Elucidating the regulation of ADAM10 is crucial for understanding the control of ADAM10-dependent cell surface proteolysis.

ADAM

Shedding

Cytoplasmic Domain

Metalloprotease

Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9)

Cell Surface Enzyme

Protein Ectodomain Shedding

Details

Title: Subtitle: The Cytoplasmic Domain of A Disintegrin and Metalloproteinase 10 (ADAM10) Regulates Its Constitutive Activity but Is Dispensable for Stimulated ADAM10-dependent Shedding
Creators: Thorsten Maretzky - From the Arthritis and Tissue Degeneration Program, Hospital for Special Surgery, New York, New York 10021
Astrid Evers - From the Arthritis and Tissue Degeneration Program, Hospital for Special Surgery, New York, New York 10021
Sylvain Le Gall - From the Arthritis and Tissue Degeneration Program, Hospital for Special Surgery, New York, New York 10021
Rolake O Alabi - From the Arthritis and Tissue Degeneration Program, Hospital for Special Surgery, New York, New York 10021
Nancy Speck - Department of Dermatology Venerology and Allergology, University Hospital Schleswig Holstein, Campus Kiel, Kiel D-24105, Germany, and the
Karina Reiss - Department of Dermatology Venerology and Allergology, University Hospital Schleswig Holstein, Campus Kiel, Kiel D-24105, Germany, and the
Carl P Blobel - From the Arthritis and Tissue Degeneration Program, Hospital for Special Surgery, New York, New York 10021
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.290(12), pp.7416-7425
DOI: 10.1074/jbc.M114.603753
PMID: 25605720
PMCID: PMC4367251
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: Elsevier Inc
Grant note: DOI: 10.13039/100000002, name: National Institutes of Health, award: R01-GM64750, T32-GM007739
Language: English
Date published: 03/20/2015
Academic Unit: Internal Medicine
Record Identifier: 9984094572802771

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