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. 1992 Nov 15;89(22):10974–10978. doi: 10.1073/pnas.89.22.10974

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.

D A Peattie 1, M W Harding 1, M A Fleming 1, M T DeCenzo 1, J A Lippke 1, D J Livingston 1, M Benasutti 1
PMCID: PMC50465  PMID: 1279700

Abstract

Using an FK506 affinity column to identify mammalian immunosuppressant-binding proteins, we identified an immunophilin with an apparent M(r) approximately 55,000, which we have named FKBP52. We used chemically determined peptide sequence and a computerized algorithm to search GenPept, the translated GenBank data base, and identified two cDNAs likely to encode the murine FKBP52 homolog. We amplified a murine cDNA fragment, used it to select a human FKBP52 (hFKBP52) cDNA clone, and then used the clone to deduce the hFKBP52 sequence (calculated M(r) 51,810) and to express hFKBP52 in Escherichia coli. Recombinant hFKBP52 has peptidyl-prolyl cis-trans isomerase activity that is inhibited by FK506 and rapamycin and an FKBP12-like consensus sequence that probably defines the immunosuppressant-binding site. FKBP52 is apparently common to several vertebrate species and associates with the 90-kDa heat shock protein (hsp90) in untransformed mammalian steroid receptor complexes. The putative immunosuppressant-binding site is probably distinct from the hsp90-binding site, and we predict that FKBP52 has different structural domains to accommodate these functions. hFKBP52 contains 12 protein kinase phosphorylation-site motifs and a potential calmodulin-binding site, implying that posttranslational phosphorylation could generate multiple isoforms of the protein and that calmodulin and intracellular Ca2+ levels could affect FKBP52 function. FKBP52 transcripts are present in a variety of human tissues and could vary in abundance and/or stability.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Altschul S. F., Lipman D. J. Protein database searches for multiple alignments. Proc Natl Acad Sci U S A. 1990 Jul;87(14):5509–5513. doi: 10.1073/pnas.87.14.5509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bresnick E. H., Dalman F. C., Pratt W. B. Direct stoichiometric evidence that the untransformed Mr 300,000, 9S, glucocorticoid receptor is a core unit derived from a larger heteromeric complex. Biochemistry. 1990 Jan 16;29(2):520–527. doi: 10.1021/bi00454a028. [DOI] [PubMed] [Google Scholar]
  3. Callebaut I., Renoir J. M., Lebeau M. C., Massol N., Burny A., Baulieu E. E., Mornon J. P. An immunophilin that binds M(r) 90,000 heat shock protein: main structural features of a mammalian p59 protein. Proc Natl Acad Sci U S A. 1992 Jul 15;89(14):6270–6274. doi: 10.1073/pnas.89.14.6270. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chou P. Y., Fasman G. D. Empirical predictions of protein conformation. Annu Rev Biochem. 1978;47:251–276. doi: 10.1146/annurev.bi.47.070178.001343. [DOI] [PubMed] [Google Scholar]
  5. Fischer G., Wittmann-Liebold B., Lang K., Kiefhaber T., Schmid F. X. Cyclophilin and peptidyl-prolyl cis-trans isomerase are probably identical proteins. Nature. 1989 Feb 2;337(6206):476–478. doi: 10.1038/337476a0. [DOI] [PubMed] [Google Scholar]
  6. Galat A., Lane W. S., Standaert R. F., Schreiber S. L. A rapamycin-selective 25-kDa immunophilin. Biochemistry. 1992 Mar 3;31(8):2427–2434. doi: 10.1021/bi00123a031. [DOI] [PubMed] [Google Scholar]
  7. Gasc J. M., Renoir J. M., Faber L. E., Delahaye F., Baulieu E. E. Nuclear localization of two steroid receptor-associated proteins, hsp90 and p59. Exp Cell Res. 1990 Feb;186(2):362–367. doi: 10.1016/0014-4827(90)90317-4. [DOI] [PubMed] [Google Scholar]
  8. Harding M. W., Galat A., Uehling D. E., Schreiber S. L. A receptor for the immunosuppressant FK506 is a cis-trans peptidyl-prolyl isomerase. Nature. 1989 Oct 26;341(6244):758–760. doi: 10.1038/341758a0. [DOI] [PubMed] [Google Scholar]
  9. Harrison R. K., Stein R. L. Substrate specificities of the peptidyl prolyl cis-trans isomerase activities of cyclophilin and FK-506 binding protein: evidence for the existence of a family of distinct enzymes. Biochemistry. 1990 Apr 24;29(16):3813–3816. doi: 10.1021/bi00468a001. [DOI] [PubMed] [Google Scholar]
  10. Hart G. W., Brew K., Grant G. A., Bradshaw R. A., Lennarz W. J. Primary structural requirements for the enzymatic formation of the N-glycosidic bond in glycoproteins. Studies with natural and synthetic peptides. J Biol Chem. 1979 Oct 10;254(19):9747–9753. [PubMed] [Google Scholar]
  11. Hart G. W., Haltiwanger R. S., Holt G. D., Kelly W. G. Glycosylation in the nucleus and cytoplasm. Annu Rev Biochem. 1989;58:841–874. doi: 10.1146/annurev.bi.58.070189.004205. [DOI] [PubMed] [Google Scholar]
  12. Heitman J., Movva N. R., Hiestand P. C., Hall M. N. FK 506-binding protein proline rotamase is a target for the immunosuppressive agent FK 506 in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1948–1952. doi: 10.1073/pnas.88.5.1948. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hung D. T., Schreiber S. L. cDNA cloning of a human 25 kDa FK506 and rapamycin binding protein. Biochem Biophys Res Commun. 1992 Apr 30;184(2):733–738. doi: 10.1016/0006-291x(92)90651-z. [DOI] [PubMed] [Google Scholar]
  14. Jin Y. J., Albers M. W., Lane W. S., Bierer B. E., Schreiber S. L., Burakoff S. J. Molecular cloning of a membrane-associated human FK506- and rapamycin-binding protein, FKBP-13. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6677–6681. doi: 10.1073/pnas.88.15.6677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jin Y. J., Burakoff S. J., Bierer B. E. Molecular cloning of a 25-kDa high affinity rapamycin binding protein, FKBP25. J Biol Chem. 1992 Jun 5;267(16):10942–10945. [PubMed] [Google Scholar]
  16. Kemp B. E., Pearson R. B. Protein kinase recognition sequence motifs. Trends Biochem Sci. 1990 Sep;15(9):342–346. doi: 10.1016/0968-0004(90)90073-k. [DOI] [PubMed] [Google Scholar]
  17. Koltin Y., Faucette L., Bergsma D. J., Levy M. A., Cafferkey R., Koser P. L., Johnson R. K., Livi G. P. Rapamycin sensitivity in Saccharomyces cerevisiae is mediated by a peptidyl-prolyl cis-trans isomerase related to human FK506-binding protein. Mol Cell Biol. 1991 Mar;11(3):1718–1723. doi: 10.1128/mcb.11.3.1718. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lane W. S., Galat A., Harding M. W., Schreiber S. L. Complete amino acid sequence of the FK506 and rapamycin binding protein, FKBP, isolated from calf thymus. J Protein Chem. 1991 Apr;10(2):151–160. doi: 10.1007/BF01024778. [DOI] [PubMed] [Google Scholar]
  19. Lebeau M. C., Massol N., Herrick J., Faber L. E., Renoir J. M., Radanyi C., Baulieu E. E. P59, an hsp 90-binding protein. Cloning and sequencing of its cDNA and preparation of a peptide-directed polyclonal antibody. J Biol Chem. 1992 Mar 5;267(7):4281–4284. [PubMed] [Google Scholar]
  20. Maki N., Sekiguchi F., Nishimaki J., Miwa K., Hayano T., Takahashi N., Suzuki M. Complementary DNA encoding the human T-cell FK506-binding protein, a peptidylprolyl cis-trans isomerase distinct from cyclophilin. Proc Natl Acad Sci U S A. 1990 Jul;87(14):5440–5443. doi: 10.1073/pnas.87.14.5440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Miyata Y., Yahara I. The 90-kDa heat shock protein, HSP90, binds and protects casein kinase II from self-aggregation and enhances its kinase activity. J Biol Chem. 1992 Apr 5;267(10):7042–7047. [PubMed] [Google Scholar]
  22. Morrison J. F. Kinetics of the reversible inhibition of enzyme-catalysed reactions by tight-binding inhibitors. Biochim Biophys Acta. 1969;185(2):269–286. doi: 10.1016/0005-2744(69)90420-3. [DOI] [PubMed] [Google Scholar]
  23. Nelson P. A., Lippke J. A., Murcko M. A., Rosborough S. L., Peattie D. A. cDNA encoding murine FK506-binding protein (FKBP): nucleotide and deduced amino acid sequence. Gene. 1991 Dec 30;109(2):255–258. doi: 10.1016/0378-1119(91)90617-k. [DOI] [PubMed] [Google Scholar]
  24. O'Neil K. T., DeGrado W. F. How calmodulin binds its targets: sequence independent recognition of amphiphilic alpha-helices. Trends Biochem Sci. 1990 Feb;15(2):59–64. doi: 10.1016/0968-0004(90)90177-d. [DOI] [PubMed] [Google Scholar]
  25. Park S. T., Aldape R. A., Futer O., DeCenzo M. T., Livingston D. J. PPIase catalysis by human FK506-binding protein proceeds through a conformational twist mechanism. J Biol Chem. 1992 Feb 15;267(5):3316–3324. [PubMed] [Google Scholar]
  26. Partaledis J. A., Fleming M. A., Harding M. W., Berlin V. Saccharomyces cerevisiae contains a homolog of human FKBP-13, a membrane-associated FK506/rapamycin binding protein. Yeast. 1992 Aug;8(8):673–680. doi: 10.1002/yea.320080812. [DOI] [PubMed] [Google Scholar]
  27. Perdew G. H., Whitelaw M. L. Evidence that the 90-kDa heat shock protein (HSP90) exists in cytosol in heteromeric complexes containing HSP70 and three other proteins with Mr of 63,000, 56,000, and 50,000. J Biol Chem. 1991 Apr 15;266(11):6708–6713. [PubMed] [Google Scholar]
  28. Renoir J. M., Radanyi C., Faber L. E., Baulieu E. E. The non-DNA-binding heterooligomeric form of mammalian steroid hormone receptors contains a hsp90-bound 59-kilodalton protein. J Biol Chem. 1990 Jun 25;265(18):10740–10745. [PubMed] [Google Scholar]
  29. Renoir J. M., Radanyi C., Jung-Testas I., Faber L. E., Baulieu E. E. The nonactivated progesterone receptor is a nuclear heterooligomer. J Biol Chem. 1990 Aug 25;265(24):14402–14406. [PubMed] [Google Scholar]
  30. Rexin M., Busch W., Gehring U. Protein components of the nonactivated glucocorticoid receptor. J Biol Chem. 1991 Dec 25;266(36):24601–24605. [PubMed] [Google Scholar]
  31. Rosborough S. L., Fleming M., Nelson P. A., Boger J., Harding M. W. Identification of FKBP-related proteins with antibodies of predetermined specificity and isolation by FK 506 affinity chromatography. Transplant Proc. 1991 Dec;23(6):2890–2893. [PubMed] [Google Scholar]
  32. Sanchez E. R., Faber L. E., Henzel W. J., Pratt W. B. The 56-59-kilodalton protein identified in untransformed steroid receptor complexes is a unique protein that exists in cytosol in a complex with both the 70- and 90-kilodalton heat shock proteins. Biochemistry. 1990 May 29;29(21):5145–5152. doi: 10.1021/bi00473a021. [DOI] [PubMed] [Google Scholar]
  33. Sanchez E. R. Hsp56: a novel heat shock protein associated with untransformed steroid receptor complexes. J Biol Chem. 1990 Dec 25;265(36):22067–22070. [PubMed] [Google Scholar]
  34. Sawada S., Suzuki G., Kawase Y., Takaku F. Novel immunosuppressive agent, FK506. In vitro effects on the cloned T cell activation. J Immunol. 1987 Sep 15;139(6):1797–1803. [PubMed] [Google Scholar]
  35. Schreiber S. L. Chemistry and biology of the immunophilins and their immunosuppressive ligands. Science. 1991 Jan 18;251(4991):283–287. doi: 10.1126/science.1702904. [DOI] [PubMed] [Google Scholar]
  36. Siekierka J. J., Hung S. H., Poe M., Lin C. S., Sigal N. H. A cytosolic binding protein for the immunosuppressant FK506 has peptidyl-prolyl isomerase activity but is distinct from cyclophilin. Nature. 1989 Oct 26;341(6244):755–757. doi: 10.1038/341755a0. [DOI] [PubMed] [Google Scholar]
  37. Smith D. F., Faber L. E., Toft D. O. Purification of unactivated progesterone receptor and identification of novel receptor-associated proteins. J Biol Chem. 1990 Mar 5;265(7):3996–4003. [PubMed] [Google Scholar]
  38. Smith R. F., Smith T. F. Pattern-induced multi-sequence alignment (PIMA) algorithm employing secondary structure-dependent gap penalties for use in comparative protein modelling. Protein Eng. 1992 Jan;5(1):35–41. doi: 10.1093/protein/5.1.35. [DOI] [PubMed] [Google Scholar]
  39. Standaert R. F., Galat A., Verdine G. L., Schreiber S. L. Molecular cloning and overexpression of the human FK506-binding protein FKBP. Nature. 1990 Aug 16;346(6285):671–674. doi: 10.1038/346671a0. [DOI] [PubMed] [Google Scholar]
  40. Tai P. K., Albers M. W., Chang H., Faber L. E., Schreiber S. L. Association of a 59-kilodalton immunophilin with the glucocorticoid receptor complex. Science. 1992 May 29;256(5061):1315–1318. doi: 10.1126/science.1376003. [DOI] [PubMed] [Google Scholar]
  41. Tai P. K., Maeda Y., Nakao K., Wakim N. G., Duhring J. L., Faber L. E. A 59-kilodalton protein associated with progestin, estrogen, androgen, and glucocorticoid receptors. Biochemistry. 1986 Sep 9;25(18):5269–5275. doi: 10.1021/bi00366a043. [DOI] [PubMed] [Google Scholar]
  42. Tocci M. J., Matkovich D. A., Collier K. A., Kwok P., Dumont F., Lin S., Degudicibus S., Siekierka J. J., Chin J., Hutchinson N. I. The immunosuppressant FK506 selectively inhibits expression of early T cell activation genes. J Immunol. 1989 Jul 15;143(2):718–726. [PubMed] [Google Scholar]
  43. Tropschug M., Wachter E., Mayer S., Schönbrunner E. R., Schmid F. X. Isolation and sequence of an FK506-binding protein from N. crassa which catalyses protein folding. Nature. 1990 Aug 16;346(6285):674–677. doi: 10.1038/346674a0. [DOI] [PubMed] [Google Scholar]
  44. Van Duyne G. D., Standaert R. F., Karplus P. A., Schreiber S. L., Clardy J. Atomic structure of FKBP-FK506, an immunophilin-immunosuppressant complex. Science. 1991 May 10;252(5007):839–842. doi: 10.1126/science.1709302. [DOI] [PubMed] [Google Scholar]
  45. Wickens M. How the messenger got its tail: addition of poly(A) in the nucleus. Trends Biochem Sci. 1990 Jul;15(7):277–281. doi: 10.1016/0968-0004(90)90054-f. [DOI] [PubMed] [Google Scholar]
  46. Wiederrecht G., Brizuela L., Elliston K., Sigal N. H., Siekierka J. J. FKB1 encodes a nonessential FK 506-binding protein in Saccharomyces cerevisiae and contains regions suggesting homology to the cyclophilins. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):1029–1033. doi: 10.1073/pnas.88.3.1029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Wiederrecht G., Martin M. M., Sigal N. H., Siekierka J. J. Isolation of a human cDNA encoding a 25 kDa FK-506 and rapamycin binding protein. Biochem Biophys Res Commun. 1992 May 29;185(1):298–303. doi: 10.1016/s0006-291x(05)80990-8. [DOI] [PubMed] [Google Scholar]
  48. Yem A. W., Tomasselli A. G., Heinrikson R. L., Zurcher-Neely H., Ruff V. A., Johnson R. A., Deibel M. R., Jr The Hsp56 component of steroid receptor complexes binds to immobilized FK506 and shows homology to FKBP-12 and FKBP-13. J Biol Chem. 1992 Feb 15;267(5):2868–2871. [PubMed] [Google Scholar]

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