Regulation of ADAM12 Cell-surface Expression by Protein Kinase C ϵ*

By Christina Sundberg, C. Thodeti, M. Kveiborg, C. Larsson, P. Parker, R. Albrechtsen, U. Wewer. 2004

71 citations recorded by the source corpus.

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) ϵ induces ADAM12 translocation to the cell surface and that catalytic activity of PKCϵ is required for this translocation. The following results support this conclusion: 1) treatment of cells with 0.1 μm phorbol 12-myristate 13-acetate (PMA) enhanced ADAM12 cell-surface immunostaining, 2) ADAM12 and PKCϵ could be co-immunoprecipitated from membrane-enriched fractions of PMA-treated cells, 3) RD cells transfected with EGFP-tagged, myristoylated PKCϵ expressed more ADAM12 at the cell surface than did non-transfected cells, and 4) RD cells transfected with a kinase-inactive PKCϵ mutant did not exhibit ADAM12 cell-surface translocation upon PMA treatment. Finally, we demonstrate that the C1 and C2 domains of PKCϵ both contain a binding site for ADAM12. These studies show that PKCϵ plays a critical role in the regulation of ADAM12 cell-surface expression.

Subfields: Molecular Biology, Oncology

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In Molecular Biology

1. M. Handsley
2. D. Edwards
3. S. Zhong
4. R. Khalil
5. C. Pennington

In Oncology

1. S. Zhong
2. M. Handsley
3. D. Edwards
4. R. Khalil
5. C. Pennington

Citations in the corpus

• A Disintegrin and Metalloproteinase (ADAM) and ADAM with Thrombospondin Motifs (ADAMTS) Family in Vascular Biology and Disease (2019, background): “Some ADAMs such as ADAM-12 are stored as already active proteins in the cell, and are transferred to the cell membrane to perform their function when the cell is simulated by cytokines [29].”
• The ADAM metalloproteinases (2008, background): “ADAM-12 is stored intracellularly as a mature, active protein but appears on the surface in response to PMA stimulation (Hougaard et al., 2000), and this regulated translocation requires the cytoplasmic domain and is dependent on PKCe (Sundberg et al., 2004).”