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

• DOI: 10.1007/s00018-015-2111-z
• Publication Date: 2015-12-19T09:58:10Z
• Authors: Stéphanie Jouannet; Julien Saint-Pol; Laurent Fernandez; Viet Nguyen; Stéphanie Charrin; Claude Boucheix; Christel Brou; Pierre-Emmanuel M...; Eric Rubinstein
• Keywords: 0301 basic medicine; Notch; Tetraspanins; Ectodomain shedding; Substrate Specificity; Cellular and Molecular Neuroscience; ADAM10 Protein; Amyloid beta-Protein Precursor; 03 medical and health sciences; Tandem Mass Spectrometry; Cell Line, Tumor; Humans; Immunoprecipitation; RNA, Small Interfering; Receptor, Notch1; Microdomain; [SDV.BC] Life Sciences [q-bio]/Cellular Biology; Molecular Biology; Chromatography, High Pressure Liquid; Pharmacology; Microscopy, Confocal; ADAM10; Membrane Proteins; Cell Biology; Tetraspanin; Cadherins; Membrane compartmentalization; ADAM Proteins; Hyaluronan Receptors; Molecular Medicine; Original Article; RNA Interference; Amyloid Precursor Protein Secretases

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.