Home LiteratureArticle Details
PMID: 8423808 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Identification of a 60-kilodalton stress-related protein, p60, which interacts with hsp90 and hsp70.

Molecular and cellular biology ·Vol. 13 ·No. 2 ·1993-02-00 ·Pages 869-76

Smith DF, Sullivan WP, Marion TN, Zaitsu K, Madden B, McCormick DJ, Toft DO

Abstract

Immunoaffinity purification of hsp90 from chick oviduct cytosol reveals two major proteins, hsp70 and a 60-kDa protein (p60), copurifying with hsp90. A similar result is obtained when hsp90 is immunoaffinity purified from chick liver and brain cytosols, avian fibroblasts, and rabbit reticulocyte lysate. This p60 is the same protein previously identified in certain assembly complexes of chick progesterone receptor generated in a cell-free reconstitution system. Tryptic and cyanogen bromide peptide fragments were generated from gel-purified p60, and partial N-terminal sequences were determined from eight peptides. The sequences show a striking similarity to the sequence of a 63-kDa human protein (IEF SSP 3521) whose abundance is increased in MRC-5 fibroblasts following simian virus 40 transformation. A monoclonal antibody was prepared against avian p60; Western immunoblot analysis showed that p60 was present in each of eight chick tissues examined and in each of the human, rat, rabbit, and Xenopus tissues tested. Immunoaffinity purifications from both chick oviduct cytosol and rabbit reticulocyte lysate using anti-p60 and anti-hsp70 monoclonal antibodies confirm that there is a relatively abundant complex in these extracts containing hsp90, hsp70, and p60. This complex appears to comprise an important functional unit in the assembly of progesterone receptor complexes. However, judging from the abundance and widespread occurrence of this multiprotein complex, hsp90, hsp70, and p60 probably function interactively in other systems as well.

MeSH Terms
Amino Acid Sequence Animals Blotting, Western Chickens Electrophoresis, Gel, Two-Dimensional Electrophoresis, Polyacrylamide Gel Heat-Shock Proteins/chemistry,metabolism Humans Molecular Chaperones Molecular Sequence Data Organ Specificity Rabbits Sequence Homology, Amino Acid Species Specificity Xenopus
Chemicals
Heat-Shock Proteins Molecular Chaperones STIP1 protein, human Stip1 protein, rat
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Smith D F
Department of Pharmacology, University of Nebraska Medical Center, Omaha 68198-6260.
Sullivan W P
Marion T N
Zaitsu K
Madden B
McCormick D J
Toft D O
References (48)
48 references, click to expand
  1. Effects of molybdate on steroid receptors in intact GH1 cells. Evidence for dissociation of an intracellular 10 S receptor oligomer prior to nuclear accumulation.
    J Biol Chem. 1985 Nov 15;260(26):14009-15 PMID: 4055769
  2. Isolation of steroid receptor binding protein from chicken oviduct and production of monoclonal antibodies.
    Biochemistry. 1985 Jul 16;24(15):4214-22 PMID: 4052391
  3. Immunological evidence that the nonhormone binding component of avian steroid receptors exists in a wide range of tissues and species.
    Biochemistry. 1985 Nov 5;24(23):6586-91 PMID: 4084541
  4. Interaction of the Rous sarcoma virus protein pp60src with the cellular proteins pp50 and pp90.
    Curr Top Microbiol Immunol. 1986;123:1-22 PMID: 2419040
  5. The common 90-kd protein component of non-transformed '8S' steroid receptors is a heat-shock protein.
    EMBO J. 1985 Dec 1;4(12):3131-5 PMID: 2419124
  6. The 70-kd mammalian heat shock proteins are structurally and functionally related to the uncoating protein that releases clathrin triskelia from coated vesicles.
    EMBO J. 1985 Dec 16;4(13A):3385-91 PMID: 2868889
  7. Uncoating ATPase is a member of the 70 kilodalton family of stress proteins.
    Cell. 1986 Apr 11;45(1):3-13 PMID: 2937542
  8. Two mammalian heat shock proteins, HSP90 and HSP100, are actin-binding proteins.
    Proc Natl Acad Sci U S A. 1986 Nov;83(21):8054-8 PMID: 3534880
  9. The 90-kilodalton peptide of the heme-regulated eIF-2 alpha kinase has sequence similarity with the 90-kilodalton heat shock protein.
    Biochemistry. 1987 Oct 20;26(21):6583-7 PMID: 3427028
  10. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa.
    Anal Biochem. 1987 Nov 1;166(2):368-79 PMID: 2449095
  11. A subfamily of stress proteins facilitates translocation of secretory and mitochondrial precursor polypeptides.
    Nature. 1988 Apr 28;332(6167):800-5 PMID: 3282178
  12. 70K heat shock related proteins stimulate protein translocation into microsomes.
    Nature. 1988 Apr 28;332(6167):805-10 PMID: 3282179
  13. Seventy-kilodalton heat shock proteins and an additional component from reticulocyte lysate stimulate import of M13 procoat protein into microsomes.
    EMBO J. 1988 Sep;7(9):2875-80 PMID: 3181144
  14. Evidence that the 90-kilodalton heat shock protein is associated with tubulin-containing complexes in L cell cytosol and in intact PtK cells.
    Mol Endocrinol. 1988 Aug;2(8):756-60 PMID: 3062385
  15. Evidence that the 90-kDa heat shock protein is necessary for the steroid binding conformation of the L cell glucocorticoid receptor.
    J Biol Chem. 1989 Mar 25;264(9):4992-7 PMID: 2647745
  16. Evidence for the association of the heme-regulated eIF-2 alpha kinase with the 90-kDa heat shock protein in rabbit reticulocyte lysate in situ.
    J Biol Chem. 1989 Sep 15;264(26):15542-7 PMID: 2768277
  17. Isolation and characterization of STI1, a stress-inducible gene from Saccharomyces cerevisiae.
    Mol Cell Biol. 1989 Sep;9(9):3638-46 PMID: 2674681
  18. Binding of heat shock proteins to the avian progesterone receptor.
    Mol Cell Biol. 1989 Sep;9(9):3829-38 PMID: 2779568
  19. Polypeptide chain binding proteins: catalysts of protein folding and related processes in cells.
    Cell. 1989 Nov 17;59(4):591-601 PMID: 2573430
  20. A repeating amino acid motif in CDC23 defines a family of proteins and a new relationship among genes required for mitosis and RNA synthesis.
    Cell. 1990 Jan 26;60(2):307-17 PMID: 2404612
  21. Purification of unactivated progesterone receptor and identification of novel receptor-associated proteins.
    J Biol Chem. 1990 Mar 5;265(7):3996-4003 PMID: 2303491
  22. Interaction of Hsp 70 with newly synthesized proteins: implications for protein folding and assembly.
    Science. 1990 May 18;248(4957):850-4 PMID: 2188360
  23. Renaturation of denatured lambda repressor requires heat shock proteins.
    Cell. 1990 Jun 15;61(6):1013-20 PMID: 2140957
  24. 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):5145-52 PMID: 2378870
  25. Structural and functional reconstitution of the glucocorticoid receptor-hsp90 complex.
    J Biol Chem. 1990 Dec 15;265(35):21397-400 PMID: 2254299
  26. Hsp56: a novel heat shock protein associated with untransformed steroid receptor complexes.
    J Biol Chem. 1990 Dec 25;265(36):22067-70 PMID: 2266108
  27. Reconstitution of progesterone receptor with heat shock proteins.
    Mol Endocrinol. 1990 Nov;4(11):1704-11 PMID: 2280772
  28. The 90-kDa heat shock protein (hsp-90) possesses an ATP binding site and autophosphorylating activity.
    J Biol Chem. 1991 Mar 15;266(8):4943-50 PMID: 2002041
  29. 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):6708-13 PMID: 2016286
  30. Phosphorylation regulates the activity of the eEF-2-specific Ca(2+)- and calmodulin-dependent protein kinase III.
    J Biol Chem. 1991 Sep 5;266(25):16425-30 PMID: 1885575
  31. A 50-kDa cytosolic protein complexed with the 90-kDa heat shock protein (hsp90) is the same protein complexed with pp60v-src hsp90 in cells transformed by the Rous sarcoma virus.
    J Biol Chem. 1991 Sep 5;266(25):16436-40 PMID: 1653236
  32. Biological role and regulation of the universally conserved heat shock proteins.
    J Biol Chem. 1991 Dec 25;266(36):24233-6 PMID: 1761528
  33. Assembly of progesterone receptor with heat shock proteins and receptor activation are ATP mediated events.
    J Biol Chem. 1992 Jan 15;267(2):1350-6 PMID: 1730655
  34. 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):2868-71 PMID: 1371107
  35. 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):4281-4 PMID: 1537818
  36. 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):7042-7 PMID: 1551911
  37. Molecular cloning and expression of a transformation-sensitive human protein containing the TPR motif and sharing identity to the stress-inducible yeast protein STI1.
    J Biol Chem. 1992 Apr 25;267(12):8485-91 PMID: 1569099
  38. Successive action of DnaK, DnaJ and GroEL along the pathway of chaperone-mediated protein folding.
    Nature. 1992 Apr 23;356(6371):683-9 PMID: 1349157
  39. Association of a 59-kilodalton immunophilin with the glucocorticoid receptor complex.
    Science. 1992 May 29;256(5061):1315-8 PMID: 1376003
  40. Hsp90 chaperones protein folding in vitro.
    Nature. 1992 Jul 9;358(6382):169-70 PMID: 1614549
  41. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  42. High resolution two-dimensional electrophoresis of proteins.
    J Biol Chem. 1975 May 25;250(10):4007-21 PMID: 236308
  43. A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines.
    J Immunol. 1979 Oct;123(4):1548-50 PMID: 113458
  44. Interaction between the Rous sarcoma virus transforming protein and two cellular phosphoproteins: analysis of the turnover and distribution of this complex.
    Mol Cell Biol. 1983 Jan;3(1):9-19 PMID: 6298609
  45. Characterization of nonactivated and activated glucocorticoid-receptor complexes from intact rat thymus cells.
    J Biol Chem. 1983 May 25;258(10):6477-85 PMID: 6853491
  46. A method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids.
    Anal Biochem. 1984 Apr;138(1):141-3 PMID: 6731838
  47. Evidence that the 90-kDa phosphoprotein associated with the untransformed L-cell glucocorticoid receptor is a murine heat shock protein.
    J Biol Chem. 1985 Oct 15;260(23):12398-401 PMID: 3900074
  48. A 90,000-dalton binding protein common to both steroid receptors and the Rous sarcoma virus transforming protein, pp60v-src.
    J Biol Chem. 1985 Nov 15;260(26):14292-6 PMID: 2414293
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.13.2.869-876.1993
Published
1993-02-00
Pages
869-76
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC358970
Subset
IM
Grants
NIDDK NIH HHS · DK 44923 · United States
NICHD NIH HHS · HD 09140 · United States
External Links
PubMed source DOI: 10.1128/mcb.13.2.869-876.1993 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2