Abstract
Two immunoglobulin production stimulating factors (IPSF) have been found in human Burkitt's lymphoma Namalwa cells. One IPSF named IPSF-IIα was purified and identified as glyceraldehyde-3-phosphate dehydrogenase as previously reported. We report here purification, identification and characterization of IPSF-IIβ. IPSF-IIβ was purified by the serial use of ammonium sulfate fractionation, hydrophobic interaction column chromatography, anion-exchange column chromatography and gel filtration. The IPSF-IIβ was estimated as a 46 KD monomeric polypeptide by gel filtration and SDS-PAGE. Partial amino acid sequence of the 46 KD protein was analyzed for 26 amino acid residues. The sequence very closely coincided with enolase (EC 4.2.1.11) derived from various origins and, it was completely homologous with that of human enolase α-chain. Rabbit muscle enolase stimulated IgM production of hybridoma lines, and IPSF-IIβ had the enzymic activity. These results suggested that IPSF-IIβ was α-enolase or its isozyme. IPSF activities of IPSF-IIβ was stable in alkaline conditions whereas the enzymic activity was rapidly lost in alkaline conditions. Though IPSF-IIβ stimulated IgM production of both human-human and mouse-mouse hybridoma lines in serum-free condition, it partially suppressed IgE production of mouse-mouse hybridoma lines.
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References
Cali L, Feo S, Oliva D and Giallongo A (1990) Nucleotide sequence of a cDNA encoding the human muscle-specific enolase (MSE). Nucl. Acids Res. 18: 1893.
Giallongo A, Feo S, Moore R, Croce CM and Showe LC (1986) Molecular cloning and nucleotide sequence of a full-length cDNA for human α enolase. Proc. Natl. Acad. Sci. USA 83: 6741–6745.
Holland MJ, Holland JP, Thill GP and Jackson KA (1981) The primary structures of two yeast enolase genes. J. Biol. Chem. 256: 1385–1395.
Iida H and Yahara I (1985) Yeast heat-shock protein of Mr 48,000 is an isoprotein of enolase. Nature 315: 688–690.
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.
Kornblatt MJ and Klugerman A (1989) Characterization of the enolase isozymes of rabbit brain: kinetic differences between mammalian and yeast enolase. Biochem. Cell. Biol. 67: 103–107.
McAleese SM, Dunbar B, Fothergill JE, Hinks LJ and Day INM (1988) Complete amino acid sequence of the neurone-specific γ isozyme of enolase (NSE) from human brain and comparison with the non-neuronal α form (NNE). Eur. J. Biochem. 178: 413–417.
Murakami H, Hashizume S, Ohashi H, Shinohara K, Yasumoto K, Nomoto K and Omura H (1985) Human-human hybridomas secreting antibodies specific to human lung carcinoma.In Vitro Cell. Develop. Biol. 21: 593–596.
Nickells RW and Browder LW (1988) A role for glyceraldehyde-3-phosphate dehydrogenase in the development of thermotorerance inXenopus laevis embryos. J. Cell Biol. 107: 1901–1909.
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. Intl. Rep. 10: 77–83.
Perucho M, Salas J and Salas ML (1980) Study of the interaction of glyceraldehyde-3-phosphate dehydrogenase with DNA. Biochim. Biophys. Acta 606: 181–195.
Russell GA, Dunbar B and Fothergill-Gilmore LA (1986) The complete amino acid sequence of chicken skeltal-muscle enolase. Biochem. J. 236: 115–126.
Ryazanov AG (1985) Glyceraldehyde-3-phosphate dehydrogenase in one of the three major RNA-binding proteins of rabbit reticulocytes. FEBS Lett. 192: 131–134.
Ryazanov AG, Ashmarina LI and Muronetz VI (1988) Association of glyceraldehyde-3-phosphate dehydrogenase with mono- and polyribosomes of rabbit reticulocytes. Eur. J. Biochem. 171: 301–305.
Sakimura K, Kushiya E, Obinata M and Takahashi Y (1985a) Molecular cloning and the nucleotide sequence of cDNA to mRNA for non-neuronal enolase (αα enolase) of rat brain and liver. Nucl. Acids Res. 13: 4365–4378.
Sakimura K, Kushiya E, Obinata M, Odani S and Takahashi Y (1985b) Molecular cloning and the nucleotide sequence of cDNA for neuron-specific enolase messenger RNA of rat brain. Proc. Natl. Acad. Sci. USA 82: 7453–7457.
Segil N, Shrutkowski A, Dworkin MB and Dworkin-Rastl E (1988) Enolase isoenzymes in adult and developingXenopus laevis and characterization of a cloned enolase sequence. Biochem. J. 251: 31–39.
Shinmoto H, Murakami H, Yamada K, Dosako S and Omura H (1988) Immunoglobulin production stimulating and inhibiting factor derived from human lung adenocarcinoma PC-8 cells. Cytotechnology 1: 295–300.
Sugahara T, Shirahata S, Yamada K and Murakami H (1991a) Purification of immunoglobulin production stimulating factor-IIα derived from Namalwa cells. Cytotechnology 5: 255–263.
Sugahara T, Shirahata S, Akiyoshi K, Isobe T, Okuyama T and Murakami H (1991b) Immunoglobulin production stimulating factor-IIα (IPSF-IIα) is glyceraldehyde-3-phosphate dehydrogenase like protein. Cytotechnology 6: 115–120.
Sugahara T, Shirahata S and Murakami H (In Preparation) The mode of actions of immunoglobulin production stimulating factor IIα (IPSF-IIα) identified as glyceraldehyde-3-phosphate dehydrogenase.
Toyoda K, Sugahara T, Inoue K, Yamada K, Shirahata S and Murakami H (1990) Purification and characterization of the immunoglobulin production stimulating factor derived from human B lymphoblastoid cell HO-323. Cytotechnology 3: 189–197.
Yamada K, Akiyoshi K, Murakami H, Sugahara T, Ikeda I, Toyoda K and Omura H (1989) Partial purification and characterization of immunoglobulin production stimulating factor derived from Namalwa cells.In vitro Cell. Develop. Biol. 25: 243–247.
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Sugahara, T., Nakajima, H., Shirahata, S. et al. Purification and characterization of immunoglobulin production stimulating factor-IIβ derived from Namalwa cells. Cytotechnology 10, 137–146 (1992). https://doi.org/10.1007/BF00570890
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DOI: https://doi.org/10.1007/BF00570890