title: FKBP12-FK506 complex inhibits phosphatase activity of two mammalian isoforms of calcineurin irrespective of their substrates or activation mechanisms.
abstract: The interaction of calcineurin (Ca2+/calmodulin-dependent protein phosphatase) with the potent immunosuppressive agent FK506 and its 12 kDa isoform binding protein (FKBP12) was investigated. The FKBP12-FK506 complex inhibited the Ca2+/calmodulin-stimulated phosphatase activity of each of two calcineurin isoforms, which contain either the catalytic subunit A alpha or A beta (calcineurin A alpha or A beta) of bovine calcineurin. Calcineurin phosphatase activity was inhibited by the FKBP12-FK506 complex irrespective of the substrate or the enzyme activation mechanism. FK506 and FKBP-12 inhibited calcineurin in a concentration-dependent manner, and complete inhibition of the phosphatase activity appeared to require a molar excess of FKBP12-FK506 complex. Immunochemical measurements revealed tissue differences in the concentration of calcineurin, which may be of importance to the selectivity for immunosuppression of all of the biological effects. Direct binding studies with [3H]dihydro-FK506 suggest that the ratio of FKBP12-FK506 complex to calcineurin in vivo when IL2 production is inhibited is well correlated with the ratio when calcineurin phosphatase activity is inhibited in vitro. These results suggest that calcineurin is a relevant cellular target of FK506 when bound to FKBP-12.
authors.affiliation: Department of Pharmacology, Kobe University School of Medicine.
authors.forename: H
authors.initials: H
authors.lastname: Mukai
authors.forename: T
authors.initials: T
authors.lastname: Kuno
authors.forename: C D
authors.initials: CD
authors.lastname: Chang
authors.forename: B
authors.initials: B
authors.lastname: Lane
authors.forename: J R
authors.initials: JR
authors.lastname: Luly
authors.forename: C
authors.initials: C
authors.lastname: Tanaka
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000201:enzymology; D019703:Calcineurin; D002148:Calmodulin-Binding Proteins / Q000032:analysis / Q000037:antagonists & inhibitors* / Q000378:metabolism; D002352:Carrier Proteins / Q000494:pharmacology*; D002413:Cations, Divalent / Q000494:pharmacology; D002417:Cattle; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D007527:Isoenzymes / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism; D008407:Mast Cells / Q000201:enzymology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000032:analysis / Q000037:antagonists & inhibitors* / Q000378:metabolism; D010766:Phosphorylation; D051381:Rats; D016559:Tacrolimus / Q000494:pharmacology*; D022021:Tacrolimus Binding Proteins
chemical_list: D002148:Calmodulin-Binding Proteins; D002352:Carrier Proteins; D002413:Cations, Divalent; D007527:Isoenzymes; D010446:Peptide Fragments; D019703:Calcineurin; D010749:Phosphoprotein Phosphatases; D022021:Tacrolimus Binding Proteins; D016559:Tacrolimus