Differential surface expression of ADAM10 and ADAM17 on human T lymphocytes and tumor cells

PLoS One. 2013 Oct 9;8(10):e76853. doi: 10.1371/journal.pone.0076853. eCollection 2013.

Abstract

A disintegrin and metalloproteases (ADAMs) have been implicated in many processes controlling organismic development and integrity. Important substrates of ADAM proteases include growth factors, cytokines and their receptors and adhesion proteins. The inducible but irreversible cleavage of their substrates alters cell-cell communication and signaling. The crucial role of ADAM proteases (e.g. ADAM10 and 17) for mammalian development became evident from respective knockout mice, that displayed pre- or perinatal lethality with severe defects in many organs and tissues. Although many substrates for these two ADAM proteases were identified over the last decade, the regulation of their surface appearance, their enzymatic activity and their substrate specificity are still not well understood. We therefore analyzed the constitutive and inducible surface expression of ADAM10 and ADAM17 on a variety of human T cell and tumor cell lines. We demonstrate that ADAM10 is constitutively present at comparably high levels on the majority of the tested cell types. Stimulation with phorbol ester and calcium ionophore does not significantly alter the amount of surface ADAM10, except for a slight down-regulation from T cell blasts. Using FasL shedding as a readout for ADAM10 activity, we show that PKC activation and calcium mobilization are both prerequisite for activation of ADAM10 resulting in a production of soluble FasL. In contrast to ADAM10, the close relative ADAM17 is detected at only low levels on unstimulated cells. ADAM17 surface expression on T cell blasts is rapidly induced by stimulation. Since this inducible mobilization of ADAM17 is sensitive to inhibitors of actin filament formation, we propose that ADAM17 but not ADAM10 is prestored in a subcellular compartment that is transported to the cell surface in an activation- and actin-dependent manner.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADAM Proteins / genetics*
  • ADAM Proteins / metabolism*
  • ADAM10 Protein
  • ADAM17 Protein
  • Actins / metabolism
  • Amyloid Precursor Protein Secretases / genetics*
  • Amyloid Precursor Protein Secretases / metabolism*
  • Cell Line, Tumor
  • Endoplasmic Reticulum / metabolism
  • Gene Expression Regulation, Neoplastic*
  • Golgi Apparatus / metabolism
  • Humans
  • Membrane Proteins / genetics*
  • Membrane Proteins / metabolism*
  • Protein Transport
  • T-Lymphocytes / cytology*
  • T-Lymphocytes / metabolism*

Substances

  • Actins
  • Membrane Proteins
  • Amyloid Precursor Protein Secretases
  • ADAM Proteins
  • ADAM10 Protein
  • ADAM10 protein, human
  • ADAM17 Protein
  • ADAM17 protein, human
  • Adam17 protein, mouse

Grant support

The work was supported by the Deutsche Forschungsgemeinschaft Bonn, Germany (SFB877, projects B4, A7) (DFG: http://www.dfg.de). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.