Neurochemical Research

, Volume 20, Issue 4, pp 497–503 | Cite as

Regulation of neuronal nitric oxide synthase by histone, protamine, and myelin basic protein

  • Jingru Hu
  • Jennifer Fridlund
  • Esam E. El-Fakahany
Original Articles


We examined the effects of endogenous basic proteins rich in the amino acidL-arginine on neuronal NO synthase activity by monitoring cyclic GMP formation in intact neuron-like neuroblastoma N1E-115 cells. Histone, protamine and myelin basic protein significantly stimulated cyclic GMP formation, both in a time- and concentration-dependent manner. These effects were blocked by hemoglobin and NO synthase inhibitors. Removal of the extracellular/intracellular Ca2+ gradient by a Ca2+ chelator completely abolished the cyclic GMP responses elicited by histone and protamine, suggesting that influex of extracellular Ca2+ might be involved in their activation of NO synthase. The effects of myelin basic protein on cyclic GMP formation, however, appeared to be due to Ca2+ release from intracellular stores. In cytosolic preparations of rat cerebellum, these basic proteins inhibited the metabolism ofL-arginine intoL-citrulline by NO synthase. We conclude from our findings that endogenous basic proteins might be involved in the regulation of neuronal NO synthase activity. Their effects on the enzyme could be either stimulatory or inhibitory, depending on whether the basic proteins exert their effects extracellularly or intracellularly, respectively.

Key Words

Basic proteins histone protamine myelin basic protein nitric oxide synthase cyclic GMP 


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  1. 1.
    Alavi, N. 1990. Effects of polycations on prostaglandin synthesis in cultured glomerular mesanglia cells. Biochim. Biophys. Acta. 1042:221–226.Google Scholar
  2. 2.
    Suzuki-Nishimura, T., Sekino, H., Yoshino, Y., Nagaya, K., Oku, N., Nango, M., and Uchida, M. K. 1989. Synthetic polycations, polyethylenimines and polyallylamines release histamine from rat mast cells. Japan. J. Pharmacol. 51:279–290.Google Scholar
  3. 3.
    Faden, A. 1992. Dynorphin increases extracellular levels of excitatory amino acids in the brain through a non-opioid mechanism. J. Neurosci. 12:425–429.Google Scholar
  4. 4.
    Hu, J., Wang, S-Z., Forray, C., and El-Fakahany, E. E. 1992. Complex allosteric modulation of cardiac muscarinic receptors by protamine: potential model for putative endogenous ligands. Mol. Pharmacol. 42:311–324.Google Scholar
  5. 5.
    Hu, J., and El-Fakahany, E. E. 1993. Allosteric interaction of dynorphin and myelin basic protein with muscarinic receptors. Pharmacology. 47:351–359.Google Scholar
  6. 6.
    Palmer, R. M. J., Ferridge, A. G., and Moncada, S. 1987. Nitric oxide accounts for the biological activity of endothelium-derived relaxing factor. Nature. 327:524–526.Google Scholar
  7. 7.
    Thomas, G., Hecker, M., and Ramwell, P. 1989. Vascular activity of polycations and basic amino acids: L-arginine does not specifically elicit endothelium-dependent relaxation. Biochem. Biophys. Res. Commun. 158:177–180.Google Scholar
  8. 8.
    Ignarro, L. J., Gold, M. E., Buga, G. M., Byrns, R. E., Wood, K. S., Chaudhuri, G., and Frank, G. 1989. Basic polyamino acids rich in arginine, lysine, or ornithine cause both enhancement of and refractoriness to formation of endothelium-derived nitric oxide in pulmonary artery and vein. Circ. Res. 64:315–329.Google Scholar
  9. 9.
    Akata, T., Yoshitaka, J-I., Nakashima, M., and Itoh, T. 1991. Effects of protamine on vascular smooth muscle of rabbit mesenteric artery. Anesthesiol. 75:833–846.Google Scholar
  10. 10.
    Moncada, S., Palmer, R. M. J. and Higgs, E. A. 1991. Nitric oxide: physiology, pathology and pharmacology. Pharmacol. Rev. 43: 109–142.Google Scholar
  11. 11.
    Snyder, S. H. 1992. Nitric oxide: first in a new class of neurotransmitters. Science 257:494–496.Google Scholar
  12. 12.
    Hecker, M., Walch, D. T. and Vane, J. R. 1991. On the substrate specificity of nitric oxide synthase. FEBS. Lett. 294:221–224.Google Scholar
  13. 13.
    Thiemermann, C., Mustafa, M., Mester, P. A., Mitchell, J.A., Hecker, M. and Vane, J.R. 1991. Inhibition of the release of endothelium-derived relaxing factor in vitro and in vivo by dipeptides containing NG-nitro-L-arginine. Br. J. Pharmacol. 104:31–38.Google Scholar
  14. 14.
    Schmidt, H. H. H. H., Lohmann, S. M. and Walter, U. 1993. The nitric oxide and cGMP signal transduction system: regulation and mechanism of action. Biochim Biophys. Acta. 1178:153–175.Google Scholar
  15. 15.
    Johns, E. W. 1964. Studies on histones. Biochem. J. 92:55–59.Google Scholar
  16. 16.
    Eylar, E. H., Brostoff, S., Hashim, G., Caccam, J., and Burnett, P. 1971. Basic A1 protein of the myelin membrane. J. Biol. Chem. 246:5770–5784.Google Scholar
  17. 17.
    Ando, T., Yamaski, M., and Sazuki, K. 1973. Isolation, characterization, structure and function. Page 1–30,in Kleinzeller A. (eds.), Protamincs, Springer Verlag, Berlin.Google Scholar
  18. 18.
    Gorsky, L. D., Förstermann, U., Ishii, K., and Murad, F. 1990. Production of an EDRF-like activity in the cytosol of N1E-15 neuroblastoma cells. FASEB J. 4:1494–1500.Google Scholar
  19. 19.
    El-Fakahany, E. E., and Richelson, E. 1980. Regulation of muscarinic receptor-mediated cyclic GMP synthesis by cultured mouse neuroblastoma cells. J. Neurochem. 35:941–948.Google Scholar
  20. 20.
    Bredt, D. S., and Snyder, S. H. 1989. Nitric oxide mediates glutamate-linked enhancement of cGMP levels in the cerebellum. Proc. Natl. Acad. Sci. USA. 86:9030–9033.Google Scholar
  21. 21.
    Hu, J., Lee, J-H., and El-Fakahany, E. E. 1994. Inhibition of neuronal nitric oxide synthase by antipsychotic drugs. Psychopharmacol. 114:161–166.Google Scholar
  22. 22.
    Lowry, O., Rosebrough, N. J., Farr, L., and Randall, R. J. 1951. Protein measurement with the folin phenol reagent. J. Biol. Chem. 193:265–275.Google Scholar
  23. 23.
    Rapoport, R. M., Ashraf, M., and Murad, F. 1989. Effects of melittin on endothelium-dependent relaxation and cyclic GMP levels in rat aorta. Circ. Res. 64:463–473.Google Scholar
  24. 24.
    Tracy, W. R., and Peach, M. J. 1993. Mechanism of mastoparan-induced EDRF release from pulmonary artery endothelial cells. J. Vasc. Res. 30:68–72.Google Scholar
  25. 25.
    Suttorp, N., Fuhrmann, M., Tannert-Otto, S., Grimminger, F., and Bhakdi, S. 1993. Pore-forming bacterial toxins potently induce release of nitric oxide in porcine endothelial cells. J. Exp. Med. 178:337–341.Google Scholar
  26. 26.
    Criscuolo, G. R., Lelkes, P. I., Rotrosen, D., and Oldfield, E. H. 1989. Cytosolic calcium changes in endothelial cells induced by a protein product of human gliomas containing vascular permeability factor activity. J. Neurosurg. 71:884–891.Google Scholar
  27. 27.
    Surichamorn, W., Forray, C., and El-Fakahany, E. E. 1990. Role of intracellular Ca2+ mobilization in muscarinic and histamine receptor-mediated activation of guanylate cyclase in N1E-115 neuroblastoma cells: assessment of the arachidonic acid release hypothesis. Mol. Pharmacol. 37:860–869.Google Scholar
  28. 28.
    Hu, J., Mahmound, M. I., and El-Fakahany, E. E. 1994. Polyamines inhibit nitric oxide synthase in rat cerebellum. Neurosci. Lett. 175: 41–45.Google Scholar
  29. 29.
    Deguchi, M., and Yoshioka, M. 1982.L-Arginine identified as an endogenous activator for soluble guanylate cyclase from neuroblastoma cells. J. Biol. Chem. 257:10147–10151.Google Scholar

Copyright information

© Plenum Publishing Corporation 1995

Authors and Affiliations

  • Jingru Hu
    • 1
  • Jennifer Fridlund
    • 1
  • Esam E. El-Fakahany
    • 1
  1. 1.Division of Neuroscience Research in PsychiatryUniversity of Minnesota School of MedicineMinneapolis

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