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Nerve growth factor binding sites on hepatic parenchymal cells

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Summary

Nerve growth factor (NGF) binding sites on rat hepatocytes (HCs) in culture for 24 to 48 h were characterized using125I-NGF. Specific binding of125I-NGF to HCs was saturable. Scatchard analysis indicated a single population of binding sites with a Kd of 5.5 nM and a Bmax of 540 fmol/mg protein. In isolated hepatocyte membranes, specific binding of125I-NGF was also apparent with Kd and Bmax values of 10.8 nM and 3740 fmol/mg protein, respectively. Specific binding of125I-NGF to HCs was displaced by excess, unlabeled NGF but not by up to 1000-fold excess of either insulin or epidermal growth factor. Internalization/sequestration of125I-NGF into HCs was measured as radioactivity present in solubilized cells after exposure to high salt and acid. These studies indicated 83±11% of125I-NGF was accumulated by internalization/sequestration at a concentration of 1 nM 125I-NGF. Internalization was reduced to 43±4% when incubations were carried out at 4° C. These results indicate the presence of a specific, low-affinity binding site for NGF on hepatocytes in culture.

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References

  • Bernd, P.; Greene, L. A. Association of125I-nerve growth factor with PC12 pheochromocytoma cells. Evidence for internalization via high-affinity receptors only and for long-term regulation by nerve growth factor of both high- and low-affinity receptors. J. Biol. Chem. 259(24):15509–15516; 1984.

    PubMed  CAS  Google Scholar 

  • Bocchini, V.; Angeletti, P. U. The nerve growth factor: purification as a 30,000-molecular-weight protein. Proc. Natl. Acad. Sci. USA 64:787–794; 1969.

    Article  PubMed  CAS  Google Scholar 

  • Bradford, M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72:248–254; 1976.

    Article  PubMed  CAS  Google Scholar 

  • Costrini, N. V.; Bradshaw, R. A. Binding characteristics and apparent molecular size of detergent-solubilized nerve growth factor receptor of sympathetic ganglia. Proc. Natl. Acad. Sci. USA 76(7):3242–3245; 1979.

    Article  PubMed  CAS  Google Scholar 

  • Ehrhard, P.; Gudat, F.; Otten, U. Measurement of human nerve growth factor binding sites in brain and in peripheral tissues by a specific immunoprecipitation assay. Naunyn-Schmiedeberg’s Arch. Pharmacol. 342:251–257; 1990.

    Article  CAS  Google Scholar 

  • Fujita, K.; Lazarovici, P.; Guroff, G. Regulation of the differentiation of PC12 pheochromocytoma cells. Environ. Health Perspective 80:127–142; 1989.

    Article  CAS  Google Scholar 

  • Guroff, G. Nerve growth factor and other factors acting on the nervous system. In: Lajtha, A., ed. Handbook of neurochemistry, vol. 5. New York: Plenum Publishing Corp.; 1983.

    Google Scholar 

  • Klaunig, J. E.; Goldblatt, P. J.; Hinton, D. E., et al. Mouse liver cell culture. I. Hepatocyte isolation. In Vitro 17(10):913–925; 1981.

    Article  PubMed  CAS  Google Scholar 

  • Koizumi, S.; Contreras, M. L.; Matsuda, Y., et al. K-252a: a specific inhibitor of the action of nerve growth factor on PC12 cells. J. Neurosci. 8(2):715–721; 1988.

    PubMed  CAS  Google Scholar 

  • Laemmli, U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680; 1970.

    Article  PubMed  CAS  Google Scholar 

  • Levi-Montalcini, R. The nerve growth factor: thirty-five years later. Science 237:1154–1162; 1987.

    Article  PubMed  CAS  Google Scholar 

  • Mazurek, N.; Weskamp, G.; Erne, P., et al. Nerve growth factor induces mast cell degranulation without changing intracellular calcium levels. FEBS. Lett. 198(2):315–320; 1986.

    Article  PubMed  CAS  Google Scholar 

  • McPherson, G. A. Kinetic, EBDA, Ligand, Lowry: a collection of radioligand binding analysis programs. Amsterdam: Elsevier Science Publishers; 1985.

    Google Scholar 

  • Otten, U.; Ehrhard, P.; Peck, R. Nerve growth factor induces growth and differentiation of human lymphocytes. Proc. Natl. Acad. Sci. USA 86:10059–10063; 1989.

    Article  PubMed  CAS  Google Scholar 

  • Savage, C. R.; Cohen, S. Epidermal growth factor and a new derivative. Rapid isolation procedures and biological and chemical characterization. J. Biol. Chem. 247(23):7609–7611; 1972.

    PubMed  CAS  Google Scholar 

  • Seglen, P. O. Preparation of rat liver cells. II. Effects of ions and chelators on tissue dispersion. Exp. Cell Res. 76:25–30; 1973a.

    Article  PubMed  CAS  Google Scholar 

  • Seglen, P. O. Preparation of rat liver cells. III. Enzymatic requirements for tissue dispersion. Exp. Cell Res. 82:391–398; 1973b.

    Article  PubMed  CAS  Google Scholar 

  • Seidl, K.; Holstein, A-F. Evidence for the presence of nerve growth factor (NGF) and NGF receptors in human testis. Cell Tissue Res. 261:549–554; 1990.

    Article  PubMed  CAS  Google Scholar 

  • Stach, R. W.; Perez-Polo, J. R. Binding of nerve growth factor to its receptor. J. Neurosci. Res. 17:1–10; 1987.

    Article  PubMed  CAS  Google Scholar 

  • Stepien, H.; Lyson, K.; Stanisz, A. M., et al. The effect of nerve growth factor on DNA synthesis, cyclic AMP and cyclic GMP accumulation by mouse spleen lymphocytes. Int. J. Immunopharmacol. 13(1):51–56; 1991.

    Article  PubMed  CAS  Google Scholar 

  • Sutter, A.; Riopelle, R. J.; Harris-Warrick, R. M., et al. Nerve growth factor receptors. Characterization of two distinct classes of binding sites on chick embryo sensory ganglia cells. J. Biol. Chem. 254:659–667; 1979.

    Google Scholar 

  • Tocco, M. D.; Contreras, M. L.; Koizumi, S., et al. Decreased levels of nerve growth factor receptor on dexamethasone-treated PC12 cells. J. Neurosci. Res. 20:411–419; 1988.

    Article  PubMed  CAS  Google Scholar 

  • Towbin, H.; Stachelin, T.; Gordon, J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA 76:4350–4354; 1979.

    Article  PubMed  CAS  Google Scholar 

  • Vaartjes, W. J.; de Haas, C. G. M.; Van den Bergh, S. G. Differential short-term effects of growth factors on fatty acid synthesis in isolated rat-liver cells. Biochem. Biophys. Res. Commun. 131(1):449–455; 1985.

    Article  PubMed  CAS  Google Scholar 

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Bailie, M.B., Roth, R.A. & Contreras, M.L. Nerve growth factor binding sites on hepatic parenchymal cells. In Vitro Cell Dev Biol - Animal 28, 621–624 (1992). https://doi.org/10.1007/BF02631037

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