Expression and characterization of human FKBP52, an immunophilin that associates with the 90-kDa heat shock protein and is a component of steroid receptor complexes.

Peattie, Debra A.; Harding, Matthew W.; Fleming, Mark; DeCenzo, Maureen T.; Lippke, Judith A.; Livingston, David J.; Benasutti, Matt

doi:10.1073/pnas.89.22.10974

Cited by 235 publications

(187 citation statements)

References 49 publications

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“…So it seems unlikely that calcineurin phosphatase inhibition is related the synergistic interaction between FK506 and steroid. Additionally, Hsp 90, hsp70 and GR were co-isolated with FKBP (Wiederrecht et al, 1992), and hsp 90 has the immunophilinbinding region (Peattie et al, 1992). It is possible that, in addition to hsp56, these proteins can interact with GR and are responsible for FK-506-mediated GR translocation.…”

Section: Discussionmentioning

confidence: 99%

FK506 Augments Glucocorticoid-Mediated Cyclooxygenase–2 Down-Regulation in Human Rheumatoid Synovial Fibroblasts

Migita

Tanaka

Okamoto

et al. 2000

Laboratory Investigation

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SUMMARY: Prostaglandins (PG) formed by cyclooxygenase (COX) enzymes are important mediators of inflammation in rheumatoid arthritis. The contribution of the inducible COX-2 to inflammation in the rheumatoid synovium is well documented. We examined the regulation of COX-2 mRNA and protein expression in response to both glucocorticoids (GC) and FK506 using rheumatoid synovial fibroblasts. Combined treatment of FK506 and a low concentration of dexamethasone (DEX) (10 Ϫ9 M) down-regulated synovial COX-2 mRNA and protein expression. In contrast, neither FK506 nor DEX (10 Ϫ9 M) alone influenced COX-2 expression. Immunocytochemical studies showed that pretreatment with FK506 enhanced the nuclear translocation of the glucocorticoid receptor (GR) in synovial fibroblasts in the presence of low concentrations of DEX (10 Ϫ9 M). Transient transfection experiments showed that treatment of cells with FK506 enhanced the expression of glucocorticoid-responsive gene reporter in the presence of DEX (10 Ϫ9 M). NF-B is known to mediate the transcriptional activation of the COX-2 gene. Electrophoretic mobility shift assay demonstrated that DNA-binding activity of NF-B was suppressed more profoundly by FK506 plus DEX (10 Ϫ9 M) treatment with those of DEX (10 Ϫ9 M) alone in IL-1␤-stimulated synovial cells. Our results indicated that FK506-induced potentiation of GR-mediated repression of synovial COX-2 gene transcription is the result of increased translocation of GR to the nucleus and subsequent repression of NF-B transactivation. Our results also suggest that FK506 may exert anti-inflammatory effects in the rheumatoid synovium by potentiating GR-mediated signal transduction. (Lab Invest 2000, 80:135-141).R heumatoid arthritis (RA) is an inflammatory joint disease in which perpetuation of synovitis leads to articular destruction (Zvaifler and Firestein, 1994). Rheumatoid synovial fibroblasts release various inflammatory mediators (Dayer et al, 1986). Prostaglandin (PG), one of the important mediators in rheumatoid synovitis, is synthesized by cyclooxygenases (COX) (Salmon et al, 1983;Wittenberg et al, 1993). It has been demonstrated that COX-2, an inducible COX isoform, is expressed in rheumatoid synovial fibroblasts and that treatment with IL-1␤ induces the synthesis of COX-2 mRNA and protein, while glucocorticoids (GC) suppress this induction (Crofford et al, 1994).GC has been used for decades as a clinical tool to suppress inflammatory and immune responses. GC action is known to be mediated by cellular glucocorticoid receptors (GR) (Bamberger et al, 1996). The binding of GC is followed by a conformational change in GR; the hormone-receptor complex translocates to the nucleus and binds to its acceptor site in DNA (Truss and Beato, 1993). In the cytoplasm, GR associates with several heat shock proteins (Pratt, 1993;Smith and Toft, 1993). Recent studies showed that the GR complex also contains members of the immunophilin class of proteins (Tai et al, 1992). Furthermore, Yem et al (Yem et al, 1992) reported that FK506-binding protein ...

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Section: Discussionmentioning

confidence: 99%

FK506 Augments Glucocorticoid-Mediated Cyclooxygenase–2 Down-Regulation in Human Rheumatoid Synovial Fibroblasts

Migita

Tanaka

Okamoto

et al. 2000

Laboratory Investigation

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“…FKBP52 displays a modular organization (11)(12)(13) and only the NH 2 -terminal part of the protein is responsible for the immunophilin character of the whole protein; it binds FK506 and rapamycin and displays a peptidylprolyl isomerase function characteristic of the immunophilin family (14). Although this NH 2 -terminal part shares high structural and functional homologies with FKBP12, it does not inhibit the phosphatase activity of calcineurin, in contrast with FKBP12 (15)(16)(17).…”

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confidence: 99%

The FKBP-associated protein FAP48 is an antiproliferative molecule and a player in T cell activation that increases IL2 synthesis

Krummrei

Baulieu

Chambraud

2003

Proc. Natl. Acad. Sci. U.S.A.

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FAP48 was identified and cloned thanks to its interaction with FK506-binding proteins (FKBPs) such as FKBP52 and FKBP12, which belong to the large family of immunophilins that bind the macrolide immunosuppressant drugs FK506 and rapamycin. We have previously shown that FAP48 -FKBP complexes are dissociated by FK506 and rapamycin, suggesting that FAP48 is an endogenous ligand of FKBP. The present work describes the biochemical consequences of FAP48 overexpression, induced by the tetracycline analogue doxycycline, in an established cell line derived from Jurkat T cells. We report that overexpression of FAP48 results in the inhibition of cellular proliferation as does the exposure of Jurkat T cells to FK506. We also show that the expression levels of argininosuccinate synthetase and the Myc antagonist Mxi1 are modified by overexpression of FAP48, suggesting that these proteins could be good candidates to mediate the antiproliferative effect of FAP48. FAP48 affects neither the calcineurin-dependent nuclear factor of activated T cells (NFAT)1 nor JNK͞p38-dependent pathways that mediate immunosuppression by FK506. However, contrary to FK506, which blocks IL2 synthesis, we observed that FAP48 -FKBP complexes increase IL2 production, thus revealing a previously uncharacterized aspect of the immunosuppressive mechanism of FK506.T he pharmacological actions of immunosuppressant drugs such as FK506 and rapamycin are mediated in the cell by intracellular proteins named FK506-binding proteins (FKBPs) belonging to the immunophilin family (reviewed in refs. 1-3).The main cytoplasmic FKBP isoform is FKBP12, which, in a complex with FK506, binds to and inhibits the phosphatase calcineurin (4-6). Subsequently, dephosphorylation and nuclear translocation of nuclear factor of activated T cells (NFAT)1 can no longer occur, leading to the inhibition of the transcription of the IL2 gene, IL2 being the major growth factor for T cells (reviewed in ref. 7). In addition, a recent report has demonstrated that immunophilin-FK506 complexes block the JNK and p38 mitogen-activated protein kinases (MAPKs) during T cell activation and also inhibit IL2 synthesis via these pathways (8). The authors showed that direct inhibitors of calcineurin activity had no effect on JNK and p38 activation, suggesting that FK506 leads to immunosuppression via calcineurin-dependent as well as calcineurin-independent pathways.FKBP52 is another member of the FKBP family that was originally found associated with heat shock protein with a molecular mass of 90 kDa (HSP90) in the heterooligomeric forms of steroid receptor complexes (9, 10). FKBP52 displays a modular organization (11-13) and only the NH 2 -terminal part of the protein is responsible for the immunophilin character of the whole protein; it binds FK506 and rapamycin and displays a peptidylprolyl isomerase function characteristic of the immunophilin family (14). Although this NH 2 -terminal part shares high structural and functional homologies with FKBP12, it does not inhibit the phosphatase activity of calc...

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“…The chaperone domain of trigger factor shows similarities with prefoldin (31)and SurA (32). FkpA (periplasmic FKBP of E. coli) (33)(34)(35) of prokaryotes and FKBP52 of eukaryotes (36) display similar combinations of prolyl isomerase and chaperone domains.…”

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confidence: 99%

Chaperone domains convert prolyl isomerases into generic catalysts of protein folding

Jakob

Žoldák

Aumüller

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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The cis/trans isomerization of peptide bonds before proline (prolyl bonds) is a rate-limiting step in many protein folding reactions, and it is used to switch between alternate functional states of folded proteins. Several prolyl isomerases of the FK506-binding protein family, such as trigger factor, SlyD, and FkpA, contain chaperone domains and are assumed to assist protein folding in vivo. The prolyl isomerase activity of FK506-binding proteins strongly depends on the nature of residue Xaa of the Xaa-Pro bond. We confirmed this in assays with a library of tetrapeptides in which position Xaa was occupied by all 20 aa. A high sequence specificity seems inconsistent with a generic function of prolyl isomerases in protein folding. Accordingly, we constructed a library of protein variants with all 20 aa at position Xaa before a rate-limiting cis proline and used it to investigate the performance of trigger factor and SlyD as catalysts of proline-limited folding. The efficiencies of both prolyl isomerases were higher than in the tetrapeptide assays, and, intriguingly, this high activity was almost independent of the nature of the residue before the proline. Apparently, the almost indiscriminate binding of the chaperone domain to the refolding protein chain overrides the inherently high sequence specificity of the prolyl isomerase site. The catalytic performance of these folding enzymes is thus determined by generic substrate recognition at the chaperone domain and efficient transfer to the active site in the prolyl isomerase domain.folding catalysis ͉ folding helpers ͉ folding mechanism ͉ SlyD ͉ trigger factor T he cis/trans isomerization of peptide bonds before proline (Xaa-Pro or prolyl bonds; Fig. 1A) is an intrinsically slow reaction that depends on the nature of the amino acid Xaa (1-3). It determines the rates of many protein folding reactions (4-6), is used as a molecular switch (7-16), and is catalyzed by prolyl isomerases (17-21). Most of the prolyl isomerases that assist in cellular protein folding contain catalytic domains that are homologous to human FKBP12 (FK-506 binding protein of 12 kDa; Fig. 1B) and chaperone domains, which interact transiently with non-native proteins (Fig. 1B). The trigger factor (22-25) and SlyD [product of the slyD (sensitive-to lysis) gene, a cytosolic Escherichia coli chaperone] (26-29) belong to this family of folding enzymes. The chaperone domain of SlyD (the ''insertin-flap'' or IF domain) shows a unique fold (30) and is structurally not related to other chaperones. The chaperone domain of trigger factor shows similarities with prefoldin (31)and SurA (32). FkpA (periplasmic FKBP of E. coli) (33-35) of prokaryotes and FKBP52 of eukaryotes (36) display similar combinations of prolyl isomerase and chaperone domains.FKBP-type prolyl isomerases are highly specific with regard to residue Xaa before the proline (37-39). An initial characterization of human FKBP12 with various proline-containing tetrapeptides and a protease-coupled assay (17) indicated that it catalyzes isomerizati...

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Expression and characterization of human FKBP52, an immunophilin that associates with the 90-kDa heat shock protein and is a component of steroid receptor complexes.

Cited by 235 publications

References 49 publications

FK506 Augments Glucocorticoid-Mediated Cyclooxygenase–2 Down-Regulation in Human Rheumatoid Synovial Fibroblasts

FK506 Augments Glucocorticoid-Mediated Cyclooxygenase–2 Down-Regulation in Human Rheumatoid Synovial Fibroblasts

The FKBP-associated protein FAP48 is an antiproliferative molecule and a player in T cell activation that increases IL2 synthesis

Chaperone domains convert prolyl isomerases into generic catalysts of protein folding

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