TspanC8 tetraspanins differentially regulate the cleavage of ADAM10 substrates, Notch activation and ADAM10 membrane compartmentalization

Cell Mol Life Sci. 2016 May;73(9):1895-915. doi: 10.1007/s00018-015-2111-z. Epub 2015 Dec 19.


The metalloprotease ADAM10 mediates the shedding of the ectodomain of various cell membrane proteins, including APP, the precursor of the amyloid peptide Aβ, and Notch receptors following ligand binding. ADAM10 associates with the members of an evolutionary conserved subgroup of tetraspanins, referred to as TspanC8, which regulate its exit from the endoplasmic reticulum. Here we show that 4 of these TspanC8 (Tspan5, Tspan14, Tspan15 and Tspan33) which positively regulate ADAM10 surface expression levels differentially impact ADAM10-dependent Notch activation and the cleavage of several ADAM10 substrates, including APP, N-cadherin and CD44. Sucrose gradient fractionation, single molecule tracking and quantitative mass-spectrometry analysis of the repertoire of molecules co-immunoprecipitated with Tspan5, Tspan15 and ADAM10 show that these two tetraspanins differentially regulate ADAM10 membrane compartmentalization. These data represent a unique example where several tetraspanins differentially regulate the function of a common partner protein through a distinct membrane compartmentalization.

Keywords: ADAM10; Ectodomain shedding; Membrane compartmentalization; Microdomain; Notch; Tetraspanin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADAM Proteins / analysis
  • ADAM Proteins / genetics
  • ADAM Proteins / metabolism*
  • ADAM10 Protein
  • Amyloid Precursor Protein Secretases / analysis
  • Amyloid Precursor Protein Secretases / genetics
  • Amyloid Precursor Protein Secretases / metabolism*
  • Amyloid beta-Protein Precursor / metabolism
  • Cadherins / metabolism
  • Cell Line, Tumor
  • Chromatography, High Pressure Liquid
  • Humans
  • Hyaluronan Receptors / metabolism
  • Immunoprecipitation
  • Membrane Proteins / analysis
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Microscopy, Confocal
  • RNA Interference
  • RNA, Small Interfering / metabolism
  • Receptor, Notch1 / genetics
  • Receptor, Notch1 / metabolism*
  • Substrate Specificity
  • Tandem Mass Spectrometry
  • Tetraspanins / antagonists & inhibitors
  • Tetraspanins / genetics
  • Tetraspanins / metabolism*


  • Amyloid beta-Protein Precursor
  • Cadherins
  • Hyaluronan Receptors
  • Membrane Proteins
  • NOTCH1 protein, human
  • RNA, Small Interfering
  • Receptor, Notch1
  • TSPAN14 protein, human
  • TSPAN15 protein, human
  • TSPAN33 protein, human
  • TSPAN5 protein, human
  • Tetraspanins
  • Amyloid Precursor Protein Secretases
  • ADAM Proteins
  • ADAM10 Protein
  • ADAM10 protein, human