TY - JOUR
T1 - The intracellular interactome of tetraspanin-enriched microdomains reveals their function as sorting machineries toward exosomes
AU - Perez-Hernandez, Daniel
AU - Gutiérrez-Vázquez, Cristina
AU - Jorge, Inmaculada
AU - López-Martín, Soraya
AU - Ursa, Angeles
AU - Sánchez-Madrid, Francisco
AU - Vázquez, Jesús
AU - Yañez-Mó, María
PY - 2013/4/26
Y1 - 2013/4/26
N2 - Extracellular vesicles are emerging as a potent mechanism of intercellular communication because they can systemically exchange genetic and protein material between cells. Tetraspanin molecules are commonly used as protein markers of extracellular vesicles, although their role in the unexplored mechanisms of cargo selection into exosomes has not been addressed. For that purpose, we have characterized the intracellular tetraspanin-enriched microdomain (TEM) interactome by high throughput mass spectrometry, in both human lymphoblasts and their derived exosomes, revealing a clear pattern of interaction networks. Proteins interacting with TEM receptors cytoplasmic regions presented a considerable degree of overlap, although some highly specific CD81 tetraspanin ligands, such as Rac GTPase, were detected. Quantitative proteomics showed that TEM ligands account for a great proportion of the exosome proteome and that a selective repertoire of CD81-associated molecules, including Rac, is not correctly routed to exosomes in cells from CD81-deficient animals. Our data provide evidence that insertion into TEM may be necessary for protein inclusion into the exosome structure.
AB - Extracellular vesicles are emerging as a potent mechanism of intercellular communication because they can systemically exchange genetic and protein material between cells. Tetraspanin molecules are commonly used as protein markers of extracellular vesicles, although their role in the unexplored mechanisms of cargo selection into exosomes has not been addressed. For that purpose, we have characterized the intracellular tetraspanin-enriched microdomain (TEM) interactome by high throughput mass spectrometry, in both human lymphoblasts and their derived exosomes, revealing a clear pattern of interaction networks. Proteins interacting with TEM receptors cytoplasmic regions presented a considerable degree of overlap, although some highly specific CD81 tetraspanin ligands, such as Rac GTPase, were detected. Quantitative proteomics showed that TEM ligands account for a great proportion of the exosome proteome and that a selective repertoire of CD81-associated molecules, including Rac, is not correctly routed to exosomes in cells from CD81-deficient animals. Our data provide evidence that insertion into TEM may be necessary for protein inclusion into the exosome structure.
UR - http://www.scopus.com/inward/record.url?scp=84876915447&partnerID=8YFLogxK
U2 - 10.1074/jbc.M112.445304
DO - 10.1074/jbc.M112.445304
M3 - Article
C2 - 23463506
AN - SCOPUS:84876915447
SN - 0021-9258
VL - 288
SP - 11649
EP - 11661
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 17
ER -