title: Hyperphosphorylation of autoantigenic targets of paraproteins is due to inactivation of PP2A.
abstract: Paratarg-7, a frequent autoantigenic target, and all other autoantigenic targets of human paraproteins molecularly defined to date are hyperphosphorylated in the respective patients compared with healthy controls, suggesting that hyperphosphorylation of autoantigenic paraprotein targets is a general mechanism underlying the pathogenesis of these paraproteins. We now show that hyperphosphorylation of paratarg-7 occurs because of an additional phosphorylation of Ser17, which is located within the paraprotein-binding epitope. Coimmunoprecipitation identified phosphokinase C ζ (PKCζ) as the kinase responsible for the phosphorylation of most, and phosphatase 2A (PP2A) as the phosphatase responsible for the dephosphorylation of all hyperphosphorylated autoantigenic targets of paraproteins. Single-nucleotide polymorphisms (SNPs) or mutations of PKCζ and PP2A were excluded. However, PP2A was inactivated by phosphorylation of its catalytic subunit at Y307. Stimulation of T cells from healthy carriers of wild-type paratarg-7 induced a partial and transient hyperphosphorylation between days 4 and 18, which was maintained by incubation with inhibitors of PP2A, again indicating that an inactivation of PP2A is responsible for the hyperphosphorylation of autoantigenic paraprotein targets. We conclude that the genetic defect underlying the dominantly inherited hyperphosphorylation of autoantigenic paraprotein targets is not in the PP2A itself, but in genes or proteins controlling PP2A activity by phosphorylation of its catalytic subunit.
authors.affiliation: José-Carreras Center for Immuno- and Gene Therapy, Department Internal Medicine I, Saarland University Medical School, Homburg (Saar), Germany.
authors.forename: Klaus-Dieter
authors.initials: KD
authors.lastname: Preuss
authors.forename: Michael
authors.initials: M
authors.lastname: Pfreundschuh
authors.forename: Natalie
authors.initials: N
authors.lastname: Fadle
authors.forename: Evi
authors.initials: E
authors.lastname: Regitz
authors.forename: Sybille
authors.initials: S
authors.lastname: Raudies
authors.forename: Niels
authors.initials: N
authors.lastname: Murwaski
authors.forename: Manfred
authors.initials: M
authors.lastname: Ahlgrimm
authors.forename: Joerg
authors.initials: J
authors.lastname: Bittenbring
authors.forename: Markus
authors.initials: M
authors.lastname: Klotz
authors.forename: Karl-Herbert
authors.initials: KH
authors.lastname: Schäfer
authors.forename: Gerhard
authors.initials: G
authors.lastname: Held
authors.forename: Frank
authors.initials: F
authors.lastname: Neumann
authors.forename: Sandra
authors.initials: S
authors.lastname: Grass
mesh_terms: D001324:Autoantigens / Q000235:genetics / Q000378:metabolism*; D001796:Blood Protein Disorders / Q000235:genetics / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D004791:Enzyme Inhibitors / Q000494:pharmacology; D000939:Epitopes / Q000276:immunology; D006801:Humans; D047468:Immunoprecipitation; D008213:Lymphocyte Activation / Q000187:drug effects / Q000276:immunology; D010266:Paraproteins / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D061251:Primary Cell Culture; D011493:Protein Kinase C / Q000378:metabolism*; D054648:Protein Phosphatase 2 / Q000037:antagonists & inhibitors / Q000378:metabolism; D021122:Protein Subunits / Q000037:antagonists & inhibitors / Q000378:metabolism; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D014162:Transfection
chemical_list: D001324:Autoantigens; D004791:Enzyme Inhibitors; D000939:Epitopes; D010266:Paraproteins; D021122:Protein Subunits; D011994:Recombinant Proteins; D011493:Protein Kinase C; D054648:Protein Phosphatase 2