title: T lymphocytes export proteasomes by way of microparticles: a possible mechanism for generation of extracellular proteasomes.
abstract: The 20S proteasome is almost exclusively localized within cells. High levels of extracellular proteasomes are also found circulating in the blood plasma of patients suffering from a variety of inflammatory, autoimmune and neoplastic diseases. However, the origin of these proteasomes remained enigmatic. Since the proteome of microparticles, small membrane enclosed vesicles released from cells, was shown to contain proteasomal subunits, we studied whether intact proteasomes are actively released into the extracellular space. Using human primary T lymphocytes stimulated with CaCl2 and the calcium ionophore A23187 to induce membrane blebbing we demonstrate that microparticles contain proteolytically active 20S proteasomes as well as the proteasome activator PA28 and subunits of the 19S proteasome regulator. Furthermore, our experiments reveal that incubation of in vitro generated T lymphocyte-microparticles with sphingomyelinase results in the hydrolysis of the microparticle membranes and subsequent release of proteasomes from the vesicles. Thus, we here show for the first time that functional proteasomes can be exported from activated immune cells by way of microparticles, the dissolution of which may finally lead to the generation of extracellular proteasomes.
authors.affiliation: Institut für Biochemie, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Isabel
authors.initials: I
authors.lastname: Bochmann
authors.forename: Frédéric
authors.initials: F
authors.lastname: Ebstein
authors.forename: Andrea
authors.initials: A
authors.lastname: Lehmann
authors.forename: Jeremias
authors.initials: J
authors.lastname: Wohlschlaeger
authors.forename: Stephan Urs
authors.initials: SU
authors.lastname: Sixt
authors.forename: Peter-Michael
authors.initials: PM
authors.lastname: Kloetzel
authors.forename: Burkhardt
authors.initials: B
authors.lastname: Dahlmann
mesh_terms: D055252:Cell-Derived Microparticles / Q000201:enzymology*; D002478:Cells, Cultured; D006801:Humans; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D021122:Protein Subunits / Q000378:metabolism; D013108:Sphingomyelin Phosphodiesterase / Q000737:chemistry; D013601:T-Lymphocytes / Q000201:enzymology / Q000378:metabolism*
chemical_list: D021122:Protein Subunits; D013108:Sphingomyelin Phosphodiesterase; D046988:Proteasome Endopeptidase Complex
keywords: T lymphocytes; circulating; extracellular; microparticles; proteasome; sphingomyelinase