Regulation of ADAM12 cell-surface expression by protein kinase C epsilon.

Christina Sundberg; Charles Kumar Thodeti; Marie Kveiborg; Christer Larsson; Peter Parker; Reidar Albrechtsen; Ulla M Wewer

doi:10.1074/jbc.M403753200

Regulation of ADAM12 cell-surface expression by protein kinase C epsilon.

Christina Sundberg, Charles Kumar Thodeti, Marie Kveiborg, Christer Larsson, Peter Parker, Reidar Albrechtsen, Ulla M Wewer

Department of Biomedical Sciences

56 Citations (Scopus)

Abstract

The ADAM (a disintegrin and metalloprotease) family consists of multidomain cell-surface proteins that have a major impact on cell behavior. These transmembrane-anchored proteins are synthesized as proforms that have (from the N terminus): a prodomain; a metalloprotease-, disintegrin-like-, cysteine-rich, epidermal growth factor-like, and transmembrane domain; and a cytoplasmic tail. The 90-kDa mature form of human ADAM12 is generated in the trans-Golgi through cleavage of the prodomain by a furin-peptidase and is stored intracellularly until translocation to the cell surface as a constitutively active protein. However, little is known about the regulation of ADAM12 cell-surface translocation. Here, we used human RD rhabdomyosarcoma cells, which express ADAM12 at the cell surface, in a temporal pattern. We report that protein kinase C (PKC) epsilon induces ADAM12 translocation to the cell surface and that catalytic activity of PKCepsilon is required for this translocation. The following results support this conclusion: 1) treatment of cells with 0.1 microM phorbol 12-myristate 13-acetate (PMA) enhanced ADAM12 cell-surface immunostaining, 2) ADAM12 and PKCepsilon could be co-immunoprecipitated from membrane-enriched fractions of PMA-treated cells, 3) RD cells transfected with EGFP-tagged, myristoylated PKCepsilon expressed more ADAM12 at the cell surface than did non-transfected cells, and 4) RD cells transfected with a kinase-inactive PKCepsilon mutant did not exhibit ADAM12 cell-surface translocation upon PMA treatment. Finally, we demonstrate that the C1 and C2 domains of PKCepsilon both contain a binding site for ADAM12. These studies show that PKCepsilon plays a critical role in the regulation of ADAM12 cell-surface expression.

Original language	English
Journal	Journal of Biological Chemistry
Volume	279
Issue number	49
Pages (from-to)	51601-11
Number of pages	10
ISSN	0021-9258
DOIs	https://doi.org/10.1074/jbc.M403753200
Publication status	Published - 2004

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@article{feb2a6605a4f11dd8d9f000ea68e967b,

title = "Regulation of ADAM12 cell-surface expression by protein kinase C epsilon.",

abstract = "The ADAM (a disintegrin and metalloprotease) family consists of multidomain cell-surface proteins that have a major impact on cell behavior. These transmembrane-anchored proteins are synthesized as proforms that have (from the N terminus): a prodomain; a metalloprotease-, disintegrin-like-, cysteine-rich, epidermal growth factor-like, and transmembrane domain; and a cytoplasmic tail. The 90-kDa mature form of human ADAM12 is generated in the trans-Golgi through cleavage of the prodomain by a furin-peptidase and is stored intracellularly until translocation to the cell surface as a constitutively active protein. However, little is known about the regulation of ADAM12 cell-surface translocation. Here, we used human RD rhabdomyosarcoma cells, which express ADAM12 at the cell surface, in a temporal pattern. We report that protein kinase C (PKC) epsilon induces ADAM12 translocation to the cell surface and that catalytic activity of PKCepsilon is required for this translocation. The following results support this conclusion: 1) treatment of cells with 0.1 microM phorbol 12-myristate 13-acetate (PMA) enhanced ADAM12 cell-surface immunostaining, 2) ADAM12 and PKCepsilon could be co-immunoprecipitated from membrane-enriched fractions of PMA-treated cells, 3) RD cells transfected with EGFP-tagged, myristoylated PKCepsilon expressed more ADAM12 at the cell surface than did non-transfected cells, and 4) RD cells transfected with a kinase-inactive PKCepsilon mutant did not exhibit ADAM12 cell-surface translocation upon PMA treatment. Finally, we demonstrate that the C1 and C2 domains of PKCepsilon both contain a binding site for ADAM12. These studies show that PKCepsilon plays a critical role in the regulation of ADAM12 cell-surface expression.",

author = "Christina Sundberg and Thodeti, {Charles Kumar} and Marie Kveiborg and Christer Larsson and Peter Parker and Reidar Albrechtsen and Wewer, {Ulla M}",

note = "Keywords: ADAM Proteins; Animals; Binding Sites; Blotting, Western; CHO Cells; Catalysis; Cell Differentiation; Cell Line, Tumor; Cell Membrane; Cricetinae; DNA, Complementary; Flow Cytometry; Gene Expression Regulation; Genetic Vectors; Golgi Apparatus; Green Fluorescent Proteins; Humans; Immunoprecipitation; Membrane Proteins; Metalloendopeptidases; Microscopy, Fluorescence; Protein Isoforms; Protein Kinase C; Protein Kinase C-epsilon; Protein Structure, Tertiary; Protein Transport; Tetradecanoylphorbol Acetate; Time Factors; Transfection",

year = "2004",

doi = "10.1074/jbc.M403753200",

language = "English",

volume = "279",

pages = "51601--11",

journal = "Journal of Biological Chemistry",

issn = "0021-9258",

publisher = "American Society for Biochemistry and Molecular Biology, Inc.",

number = "49",

}

TY - JOUR

T1 - Regulation of ADAM12 cell-surface expression by protein kinase C epsilon.

AU - Sundberg, Christina

AU - Thodeti, Charles Kumar

AU - Kveiborg, Marie

AU - Larsson, Christer

AU - Parker, Peter

AU - Albrechtsen, Reidar

AU - Wewer, Ulla M

N1 - Keywords: ADAM Proteins; Animals; Binding Sites; Blotting, Western; CHO Cells; Catalysis; Cell Differentiation; Cell Line, Tumor; Cell Membrane; Cricetinae; DNA, Complementary; Flow Cytometry; Gene Expression Regulation; Genetic Vectors; Golgi Apparatus; Green Fluorescent Proteins; Humans; Immunoprecipitation; Membrane Proteins; Metalloendopeptidases; Microscopy, Fluorescence; Protein Isoforms; Protein Kinase C; Protein Kinase C-epsilon; Protein Structure, Tertiary; Protein Transport; Tetradecanoylphorbol Acetate; Time Factors; Transfection

PY - 2004

Y1 - 2004

N2 - The ADAM (a disintegrin and metalloprotease) family consists of multidomain cell-surface proteins that have a major impact on cell behavior. These transmembrane-anchored proteins are synthesized as proforms that have (from the N terminus): a prodomain; a metalloprotease-, disintegrin-like-, cysteine-rich, epidermal growth factor-like, and transmembrane domain; and a cytoplasmic tail. The 90-kDa mature form of human ADAM12 is generated in the trans-Golgi through cleavage of the prodomain by a furin-peptidase and is stored intracellularly until translocation to the cell surface as a constitutively active protein. However, little is known about the regulation of ADAM12 cell-surface translocation. Here, we used human RD rhabdomyosarcoma cells, which express ADAM12 at the cell surface, in a temporal pattern. We report that protein kinase C (PKC) epsilon induces ADAM12 translocation to the cell surface and that catalytic activity of PKCepsilon is required for this translocation. The following results support this conclusion: 1) treatment of cells with 0.1 microM phorbol 12-myristate 13-acetate (PMA) enhanced ADAM12 cell-surface immunostaining, 2) ADAM12 and PKCepsilon could be co-immunoprecipitated from membrane-enriched fractions of PMA-treated cells, 3) RD cells transfected with EGFP-tagged, myristoylated PKCepsilon expressed more ADAM12 at the cell surface than did non-transfected cells, and 4) RD cells transfected with a kinase-inactive PKCepsilon mutant did not exhibit ADAM12 cell-surface translocation upon PMA treatment. Finally, we demonstrate that the C1 and C2 domains of PKCepsilon both contain a binding site for ADAM12. These studies show that PKCepsilon plays a critical role in the regulation of ADAM12 cell-surface expression.

AB - The ADAM (a disintegrin and metalloprotease) family consists of multidomain cell-surface proteins that have a major impact on cell behavior. These transmembrane-anchored proteins are synthesized as proforms that have (from the N terminus): a prodomain; a metalloprotease-, disintegrin-like-, cysteine-rich, epidermal growth factor-like, and transmembrane domain; and a cytoplasmic tail. The 90-kDa mature form of human ADAM12 is generated in the trans-Golgi through cleavage of the prodomain by a furin-peptidase and is stored intracellularly until translocation to the cell surface as a constitutively active protein. However, little is known about the regulation of ADAM12 cell-surface translocation. Here, we used human RD rhabdomyosarcoma cells, which express ADAM12 at the cell surface, in a temporal pattern. We report that protein kinase C (PKC) epsilon induces ADAM12 translocation to the cell surface and that catalytic activity of PKCepsilon is required for this translocation. The following results support this conclusion: 1) treatment of cells with 0.1 microM phorbol 12-myristate 13-acetate (PMA) enhanced ADAM12 cell-surface immunostaining, 2) ADAM12 and PKCepsilon could be co-immunoprecipitated from membrane-enriched fractions of PMA-treated cells, 3) RD cells transfected with EGFP-tagged, myristoylated PKCepsilon expressed more ADAM12 at the cell surface than did non-transfected cells, and 4) RD cells transfected with a kinase-inactive PKCepsilon mutant did not exhibit ADAM12 cell-surface translocation upon PMA treatment. Finally, we demonstrate that the C1 and C2 domains of PKCepsilon both contain a binding site for ADAM12. These studies show that PKCepsilon plays a critical role in the regulation of ADAM12 cell-surface expression.

U2 - 10.1074/jbc.M403753200

DO - 10.1074/jbc.M403753200

M3 - Journal article

C2 - 15364951

SN - 0021-9258

VL - 279

SP - 51601

EP - 51611

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 49

ER -

Regulation of ADAM12 cell-surface expression by protein kinase C epsilon.

Abstract

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