scholarly journals The 59 kDa FK506-binding protein, a 90 kDa heat shock protein binding immunophilin (FKBP59-HBI), is associated with the nucleus, the cytoskeleton and mitotic apparatus

1995 ◽  
Vol 108 (5) ◽  
pp. 2037-2051 ◽  
Author(s):  
M. Perrot-Applanat ◽  
C. Cibert ◽  
G. Geraud ◽  
J.M. Renoir ◽  
E.E. Baulieu
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Protein Binding ◽  
Steroid Receptors ◽  
Binding Protein ◽  
Lymphoid Cells ◽  
Confocal Laser ◽  
Mitotic Apparatus ◽  
Fk506 Binding Protein

FKBP59-HBI, a 59 kDa FK506 binding protein which binds the 90 kDa heat shock protein hsp90 and thus is a heat shock protein binding immunophilin (HBI), was originally discovered in association with unliganded steroid receptors in their heat shock protein containing heterooligomer form. It belongs to a growing family including other FKBPs which bind the immunosuppressants FK506 and rapamycin, and cyclophilins which bind cyclosporin A, all having rotamase (peptidyl-prolyl cis-trans isomerase) activity which may be involved in protein folding. Targets for drug-immunophilin complexes have been mostly studied in vivo in T lymphocytes; however, immunophilins are present in all cell types, where their role and distribution are still unknown. Here we report the localization of FKBP59-HBI in various non lymphoid cells (mouse fibroblasts (L-929), monkey kidney cells (Cos-7), Madin-Darby canine kidney epithelial cells (MDCK), and mouse neuronal cells (GT1)). Two polyclonal antipeptide antibodies directed against the C-terminal end (amino acids 441–458) (Ab 173) or the sequence 182–201 (Ab 790) of the FKBP59-HBI were used in light and confocal laser immunofluorescence. FKBP59-HBI was found in the cytoplasm and nucleus of interphase cells. Specific immunofluorescence was much stronger in the cytoplasm than in the nucleus when using Ab 173, and stronger in the nucleus than in the cytoplasm with Ab 790. Detailed observations of L-cells, which have a particularly flat morphology, showed a punctate as well as a fibrous cytoskeletal staining in the cytoplasm using antibody 173, a result which suggests interactions of FKBP59-HBI with an organized network. Colocalization experiments (using antibodies against tubulin, vimentin or actin) and use of cytoskeletal-disrupting drugs revealed partial association of FKBP59-HBI with the microtubules. Western blot experiments confirmed that the protein was present in the subcellular fractions containing either ‘soluble’ proteins released from cells exposed to NP40 detergent, or proteins released from the cytoskeleton exposed to calcium ions (i.e. in microtubule depolymerizing conditions). Exposure of cells to 1 microM FK506 and rapamycin for 1 hour did not modify significantly the staining, although rapamycin treatment rendered the network stained by 173 clearly visible. Interestingly, during mitosis FKBP59-HBI segregated from the region of the chromosomes; it mainly localized with the mitotic apparatus (centrosome, spindle and interzone separating the chromosomes), the cleavage furrow and the midbodies during cytokinesis. It appeared again as a fibrous network in the cytoplasm of the two daughters cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Rabbit FKBP59-heat shock protein binding immunophillin (HBI) is a calmodulin binding protein

1992 ◽  
Vol 187 (3) ◽  
pp. 1330-1335 ◽  
Author(s):  
Nelly Massol ◽  
Marie-Claire Lebeau ◽  
Jack-Michel Renoir ◽  
Lee E. Faber ◽  
Etienne-Emile Baulieu
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Protein Binding ◽  
Binding Protein ◽  
Calmodulin Binding

The Mammalian Heat Shock Protein Binding Immunophilin (p59/HBI) Is an ATP and GTP Binding Protein

1993 ◽  
Vol 195 (2) ◽  
pp. 600-607 ◽  
Author(s):  
S. Lebihan ◽  
J.M. Renoir ◽  
C. Radanyi ◽  
B. Chambraud ◽  
V. Joulin ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Protein Binding ◽  
Binding Protein ◽  
Gtp Binding Protein ◽  
Gtp Binding

A constitutive 70 kDa heat-shock protein is localized on the fibres of spindles and asters at metaphase in an ATP-dependent manner: a new chaperone role is proposed

2001 ◽  
Vol 360 (2) ◽  
pp. 413-419 ◽  
Author(s):  
Cecilia AGUELI ◽  
Fabiana GERACI ◽  
Giovanni GIUDICE ◽  
Laura CHIMENTI ◽  
Domenico CASCINO ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Paracentrotus Lividus ◽  
Immunoblot Analysis ◽  
Dependent Manner ◽  
Mitotic Apparatus ◽  
T Complex ◽  
A Subunit ◽  
Cell Division Control

In the present study, double immunofluorescence and immunoblot analysis have been used to show that centrosomes, isolated from Paracentrotus lividus sea urchin embryos at the first mitotic metaphase, contain the constitutive chaperone, heat-shock protein (HSP) 70. More specifically, we demonstrate that centrosomes contain only the HSP70-d isoform, which is one of the four isoforms identified in P. lividus. We also provide evidence that p34(cell division control kinase-2) and t complex polypeptide-1 (TCP-1) α, a subunit of the TCP-1 complex, are localized on the centrosomes. Furthermore, inhibition of TCP-1 in vivo, via microinjecting an anti-(TCP-1α) antibody into P. lividus eggs before fertilization, either impaired mitosis or induced severe malformations in more than 50% of embryos. In addition, we have isolated the whole mitotic apparatus and shown that HSP70 localizes on the fibres of spindles and asters, and binds them in an ATP-dependent manner. These observations suggest that HSP70has a chaperone role in assisting the TCP-1 complex in tubulin folding, when localized on centrosomes, and during the assembling and disassembling of the mitotic apparatus, when localized on the fibres of spindles and asters.

scholarly journals Distinct functions of the 90 kDa heat-shock protein (hsp90) in oestrogen and mineralocorticosteroid receptor activity: effects of hsp90 deletion mutants

1995 ◽  
Vol 311 (3) ◽  
pp. 797-804 ◽  
Author(s):  
N Binart ◽  
M Lombès ◽  
E E Baulieu
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Leucine Zipper ◽  
Nuclear Translocation ◽  
Steroid Receptors ◽  
Binding Capacity ◽  
Hormone Binding ◽  
Affinity Ligand

Recent studies have confirmed that the 90 kDa heat-shock protein (hsp90) interacts both in vitro and in vivo with steroid receptors, encouraging further detailed physicochemical and functional analysis of its chaperone role. Thus, to explore the relationship between hsp90 and receptors, the baculovirus system was used to overexpress the chick hsp90 alpha (chsp90) along with the chick oestradiol receptor (cER) or the human mineralocorticosteroid receptor (hMR). These receptors were able to form 9 S complexes with chsp90, demonstrating the association of the co-expressed recombinant proteins. Three mutants of chsp90 (delta A, delta B and delta Z) have been created by deletion of the A (residues 221-290) and B (530-581) regions, rich in charged amino acids, and the Z (392-419) region, a putative leucine zipper. After co-expression, anti-receptor antibodies immunoprecipitated the cER or hMR complexed with the wild-type chsp90, the delta B or the delta Z mutant, but not with the delta A chsp90, indicating that deletion of the A region of chsp90 leads to a lack of interaction with these receptors. The hormone binding capacity of the cER was unaffected after its co-expression with each of the three mutants. In contrast, the hMR co-expressed with the delta B mutant failed to bind aldosterone, a finding confirmed in vivo by the absence of hormone-induced hMR nuclear translocation. Thus the B region is required for high-affinity ligand binding by the hMR. Our results suggest that the A region (but not the B or Z regions) is involved in binding of chsp90 to the cER and hMR, while the B region is essential for hormone binding by the hMR, consistent with a chaperone function for hsp90.

P59 (FK506 binding protein 59) interaction with heat shock proteins is highly conserved and may involve proteins other than steroid receptors

Biochemistry ◽  
1993 ◽  
Vol 32 (34) ◽  
pp. 8842-8847 ◽  
Author(s):  
Ping Kaung K. Tai ◽  
Hong Chang ◽  
Mark W. Albers ◽  
Stuart L. Schreiber ◽  
David O. Toft ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Proteins ◽  
Steroid Receptors ◽  
Binding Protein ◽  
Fk506 Binding Protein ◽  
Shock Proteins

Development of a sensitive assay for the measurement of antibodies against heat shock protein binding protein 1 (HspBP1): Increased levels of anti-HspBP1 IgG are prevalent in HIV infected subjects

2005 ◽  
Vol 76 (4) ◽  
pp. 464-469 ◽  
Author(s):  
Diana Papp ◽  
Zoltán Prohászka ◽  
Judit Kocsis ◽  
George Füst ◽  
Dénes Bánhegyi ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Protein Binding ◽  
Binding Protein ◽  
Sensitive Assay

scholarly journals Structure-Function Analysis of Squirrel Monkey FK506-Binding Protein 51, a Potent Inhibitor of Glucocorticoid Receptor Activity

Endocrinology ◽  
2005 ◽  
Vol 146 (7) ◽  
pp. 3194-3201 ◽  
Author(s):  
Wesley B. Denny ◽  
Viravan Prapapanich ◽  
David F. Smith ◽  
Jonathan G. Scammell
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Glucocorticoid Receptor ◽  
Squirrel Monkey ◽  
Inhibitory Activity ◽  
Function Analysis ◽  
Binding Protein ◽  
Heat Shock Protein 90 ◽  
Ppiase Activity ◽  
Fk506 Binding Protein

Abstract FK506-binding protein 51 (FKBP51) and FKBP52 are large molecular weight immunophilins that are part of the mature glucocorticoid receptor (GR) heterocomplex. These proteins possess peptidyl-prolyl isomerase (PPIase) and tetratricopeptide repeats (TPR) domains that are important for modulation of GR activity. A naturally occurring animal model of glucocorticoid resistance, the squirrel monkey, results from the relative overexpression of FKBP51 that renders the GR in a low-affinity state. In vitro studies demonstrated that the squirrel monkey form of FKBP51 is greater than 6-fold more potent than human FKBP51 in this respect. The goals of these studies were to determine the roles of the TPR and PPIase domains in the inhibitory activity of squirrel monkey FKBP51 and to gain insight into structural features of squirrel monkey FKBP51 responsible for potent inhibition of dexamethasone-stimulated GR activity. Mutations in the TPR of squirrel monkey FKBP51 that inhibit association with heat shock protein 90 blocked GR inhibitory activity. Mutations that abrogate the PPIase activity of squirrel monkey FKBP51 had no effect on GR inhibitory activity. Chimeras of squirrel monkey and human FKBP51 were tested to identify domains responsible for their different inhibitory potencies. Amino acid differences in domains FK1 and FK2 between squirrel monkey and human FKBP51 contribute equally to the enhanced inhibitory activity of squirrel monkey FKBP51. Furthermore, squirrel monkey FKBP51 in which either FK1 or FK2 was deleted lacked GR inhibitory activity. Thus, the potent inhibitory activity of squirrel monkey FKBP51 involves both FK domains and the heat shock protein 90-binding TPR domain.

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