Skip to main content
Log in

Growth stimulating activity of heat shock protein 90α to lymphoid cell lines in serum-free medium

  • Original Research Papers
  • Published:
Cytotechnology Aims and scope Submit manuscript

Abstract

A growth stimulating factor was purified from the culture supernatant of human-human hybridoma SH-76 cells in serum-free RPMI 1640 medium by the serial use of DEAE anion and heparin affinity chromatographies. The factor was a 85 kDa protein on SDS-polyacrylamide gel electrophoresis. N-terminal amino acid sequence (PEETQTQDQPME) of the protein was coincident with that of heat shock protein 90α (HSP90α). The protein reacted with anti-HSP90 monoclonal antibody. These results suggest that the protein was a member of HSP90α family after taking other circumstantial evidence into account. The protein stimulated the growth of some lymphoid cell lines such as human B-lymphoblastoid cell line HO-323-3 hybridomas derived from HO-323, and several other lymphoid cell lines. There were several lymphoid cell lines which did not respond to the protein. Growth stimulating activity of the protein was heat-unstable and significantly decreased at above 60°C. These are the first data that describe growth-stimulating activity of heat shock protein 90α.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Brugge JS, Erikson E and Erikson RL (1981) The specific interaction of the Rous sarcoma virus transforming protein, pp60src, with two cellular proteins, Cell 25: 363–372.

    Article  CAS  Google Scholar 

  • Hashizume S, Murakami H, Hirose S, Shirai T, Shinohara K and Omura H (1986) Human-human hybridomas from lung cancer patients secrete anti-DNA antibodies. Agric. Biol. Chem. 50: 1351–1353.

    CAS  Google Scholar 

  • Kamei Y, Potey JL, Yoshimizu M, Kimura T, Yamada K, Shirahata S and Murakami H (1990) Antigenic analysis of fish viruses with monoclonal antibodies. In: Trends in Animal Cell Culture Technology, pp. 201–204.

  • Kawahara H, Shirahata S, Tachibana H and Murakami H (1992)In vitro immunization of human lymphocytes with human lung cancer cell line A549. Hum. Antibod. Hybridomas 3: 8–13.

    CAS  Google Scholar 

  • Koyama K, Akiyama K, Kawahara H, Egashira A and Murakami H (1990) Alloimmunization with cultured human stomach cancer cell lines and the establishment of human-human hybridomas producing monoclonal antibodies. Jpn. J. Cancer Res. 81: 967–970.

    CAS  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural protein during assembly of head of bacteriophage-T4. Nature 227: 680–685.

    Article  CAS  Google Scholar 

  • Mendel DB and Orti E (1988) Isoform composition and stoichiometry of 90-kDa heat shock protein associated with glucocorticoid receptors. J. Biol. Chem. 263: 6695–6702.

    CAS  Google Scholar 

  • Miyata M, Murakami H, Hashizume S, Yamada K and Ohmura H (1988) Purification and characterization of lymphocytic clonal growth factor (LCGF) derived from human-human hybridoma SH-76 cells. Cytotechnology 1: 347–353.

    Article  Google Scholar 

  • Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxity assays. J. Immunol. Methods 65: 55–63.

    Article  CAS  Google Scholar 

  • Murakami H (1989) Serum-free media used for cultivation of hybridomas. In: Advances in Biotechnological Press, vol. 11, Alan R. Liss Inc., pp. 107–141.

  • Murakami H, Hashizume S and Ohashi H (1985) Human-human hybridomas secreting antibodies specific to human lung carcinoma.In vitro Cell Develop. Biol. 21: 593–596.

    CAS  Google Scholar 

  • Murakami H, Masui H, Sato GH, Sueoka N, Chow TP and Kano-Sueoka T (1982) Growth of hybridoma cells in serum-free medium: ethanolamine is an essential component. Proc. Natl. Acad. Sci. U.S.A. 79: 1158–1162.

    Article  CAS  Google Scholar 

  • Oppermann H, Levinson W and Bishop JM (1981) A cellular protein that associates with the transforming protein of Rous sarcoma virus is also a heat-shock protein. Proc. Natl. Acad. Sci. U.S.A. 78: 1067–1071.

    Article  CAS  Google Scholar 

  • Ohashi H, Hashizume S, Murakami H, Aihara K, Shinohara K and Omura H (1986) HO-323, a human B-lymphoblastoid cell line useful for making human-human hybridomas. Cell Biol. Intern. Reports 10: 77–83.

    Article  CAS  Google Scholar 

  • Rebbe NF, Ware J, Bertina RM, Modrich P and Stafford DW (1987) Nucleotide sequence of a cDNa for a member of the human 90-kDa heat-shock protein family. Gene 53: 235–245.

    Article  CAS  Google Scholar 

  • Rose DW, Wettenhall REH, Kudlicki W, Kramer G and Hardesty B (1987) The 90-kilodalton peptide of the heme-regulated eIF-2α kinase has sequence similarity with the 90-kilodalton heat shock protein. J. Biol. Chem. 26: 6583–6587.

    CAS  Google Scholar 

  • Sanchez ER, Yoft DO, Schlesinger MJ and Pratt WB (1985) Evidence that the 90-kDa phosphoprotein associated with the untransformed L-cell glucocorticoid receptor is a murine heat shock protein. J. Biol. Chem. 260: 12398–12401.

    CAS  Google Scholar 

  • Simon MC, Kitchner K, Kao H-T, Hickey E, Weber L, Voellmy R, Heintz N and Nevins JR (1987) Selective induction of human heat-shock gene transcription by the adenovirus EIA gene products, including the 12 S E1A product. Mol. Cell. Biol. 7: 2884–2890.

    CAS  Google Scholar 

  • Yahara I, Iida H and Koyasu S (1986) A heat shock-resistant variant of Chinese hamster cell line constitutively expressing heat shock protein of Mr 90,000 at high level. Cell Struct. Funct. 11: 65–73.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kuroita, T., Tachibana, H., Ohashi, H. et al. Growth stimulating activity of heat shock protein 90α to lymphoid cell lines in serum-free medium. Cytotechnology 8, 109–117 (1992). https://doi.org/10.1007/BF02525493

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02525493

Key words

Navigation