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. 1982 Jun 1;93(3):638–647. doi: 10.1083/jcb.93.3.638

Serotonin storage pools in basophil leukemia and mast cells: characterization of two types of serotonin binding protein and radioautographic analysis of the intracellular distribution of [3H]serotonin

PMCID: PMC2112151  PMID: 7118996

Abstract

We studied binding of serotonin to protein(s) derived from rat basophil leukemia (RBL) cells and mast cells. We found two types of serotonin binding protein in RBL cells. These proteins differed from one another in molecular weight and eluted in separate peaks from sephadex G-200 columns. Peak I protein (KD = 1.9 X 10(-6) M) was a glycoprotein that bound to concanavalin A (Con A); Peak II protein (KD1 = 4.5 X 10(-8) M; KD2 = 3.9 X 10(-6) M) did not bind to Con A. Moreover, binding of [3H]serotonin to protein of peak I was sensitive to inhibition by reserpine, while binding of [3H]serotonin to protein of peak II resisted inhibition by that drug. Other differences between the two types of binding protein were found, the most significant of which was the far more vigorous conditions of homogenization required to extract peak I than peak II protein. Neither peak I nor peak II protein resembled the serotonin binding protein (SBP) that is found in serotonergic neurons of the brain and gut. Electron microscope radioautographic analysis of the intracellular distribution of [3H]serotonin taken up in vitro by RBL cells or in vivo by murine mast cells indicated that essentially all of the labeled amine was located in cytoplasmic granules. No evidence for a pool in the cytosol was found and all granules were capable of becoming labeled. The presence of two types of intracellular serotonin binding proteins in these cells may indicate that there are two intracellular storage compartments for the amine. Both may be intragranular, but peak I protein may be associated with the granular membrane while peak II protein may be more free within the granular core. Different storage proteins may help to explain the differential release of amines from mast cell granules.

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Selected References

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  1. Askenase P. W., Bursztajn S., Gershon M. D., Gershon R. K. T cell-dependent mast cell degranulation and release of serotonin in murine delayed-type hypersensitivity. J Exp Med. 1980 Nov 1;152(5):1358–1374. doi: 10.1084/jem.152.5.1358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BENDITT E. P., WONG R. L., ARASE M., ROEPER E. 5-Hydroxytryptamine in mast cells. Proc Soc Exp Biol Med. 1955 Oct;90(1):303–304. doi: 10.3181/00379727-90-22016. [DOI] [PubMed] [Google Scholar]
  3. Bernd P., Gershon M. D., Nunez E. A., Tamir H. Separation of dissociated thyroid follicular and parafollicular cells: association of serotonin binding protein with parafollicular cells. J Cell Biol. 1981 Mar;88(3):499–508. doi: 10.1083/jcb.88.3.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Da Prada M., Pletscher A. Isolated 5-hydroxytryptamine organelles of rabbit blood platelets: physiological properties and drug-induced changes. Br J Pharmacol. 1968 Nov;34(3):591–597. doi: 10.1111/j.1476-5381.1968.tb08487.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  6. Falck B., Owman C. 5-hydroxytryptamine and related amines in endocrine cell systems. Adv Pharmacol. 1968;6(Pt A):211–231. doi: 10.1016/s1054-3589(08)61175-8. [DOI] [PubMed] [Google Scholar]
  7. Fillion G., Beaudoin D., Rousselle J. C., Jacob J. [3H]5-HT binding sites and 5-HT-sensitive adenylate cyclase in glial cell membrane fraction. Brain Res. 1980 Oct 6;198(2):361–374. doi: 10.1016/0006-8993(80)90750-7. [DOI] [PubMed] [Google Scholar]
  8. Gershon M. D., Nunez E. A. Histochemical and radioautographic studies of serotonin and parafollicular cells in the thyroid gland of the prehibernating bat. Endocrinology. 1970 Jan;86(1):160–166. doi: 10.1210/endo-86-1-160. [DOI] [PubMed] [Google Scholar]
  9. Gershon M. D., Nunez E. A. Uptake of biogenic amines by thyroid glands. Mol Cell Endocrinol. 1976 Aug-Sep;5(3-4):169–180. doi: 10.1016/0303-7207(76)90081-2. [DOI] [PubMed] [Google Scholar]
  10. Gershon M. D., Tamir H. Serotonin binding protein: role in transmitter storage in central and peripheral serotonergic neurons. Adv Exp Med Biol. 1981;133:37–49. doi: 10.1007/978-1-4684-3860-4_2. [DOI] [PubMed] [Google Scholar]
  11. Gershon M. D. The enteric nervous system. Annu Rev Neurosci. 1981;4:227–272. doi: 10.1146/annurev.ne.04.030181.001303. [DOI] [PubMed] [Google Scholar]
  12. Gershon R. K., Askenase P. W., Gershon M. D. Requirement for vasoactive amines for production of delayed-type hypersensitvity skin reactions. J Exp Med. 1975 Sep 1;142(3):732–747. doi: 10.1084/jem.142.3.732. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hertz L., Baldwin F., Schousboe A. Serotonin receptors on astrocytes in primary cultures: effects of methysergide and fluoxetine. Can J Physiol Pharmacol. 1979 Feb;57(2):223–226. doi: 10.1139/y79-034. [DOI] [PubMed] [Google Scholar]
  14. Isersky C., Metzger H., Buell D. N. Cell cycle-associated changes in receptors for IgE during growth and differentiation of a rat basophilic leukemia cell line. J Exp Med. 1975 May 1;141(5):1147–1162. doi: 10.1084/jem.141.5.1147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jaim-Etcheverry G., Zieher L. M. Cytochemical localization of monoamine stores in sheep thyroid gland at the electron microscope level. Experientia. 1968 Jun 15;24(6):593–595. doi: 10.1007/BF02153794. [DOI] [PubMed] [Google Scholar]
  16. Jonakait G. M., Tamir H., Gintzler A. R., Gershon M. D. Release of [3H]serotonin and its binding protein from enteric neurons. Brain Res. 1979 Sep 28;174(1):55–69. doi: 10.1016/0006-8993(79)90803-5. [DOI] [PubMed] [Google Scholar]
  17. Mahadik S. P., Korenovsky A., Rapport M. M. Slab gel analysis of the polypeptide components of rat brain subcellular organelles. Anal Biochem. 1976 Dec;76(2):615–633. doi: 10.1016/0003-2697(76)90355-9. [DOI] [PubMed] [Google Scholar]
  18. Morrison D. C., Roser J. F., Henson P. M., Cochrane C. G. Activation of rat mast cells by low molecular weight stimuli. J Immunol. 1974 Feb;112(2):573–582. [PubMed] [Google Scholar]
  19. Tamir H., Bebirian R., Muller F., Casper D. Differences between intracellular platelet and brain proteins that bind serotonin. J Neurochem. 1980 Nov;35(5):1033–1044. doi: 10.1111/j.1471-4159.1980.tb07857.x. [DOI] [PubMed] [Google Scholar]
  20. Tamir H., Brunner W., Casper D., Rapport M. M. Enhancement by gangliosides of the binding of serotonin to serotonin binding protein. J Neurochem. 1980 Jun;34(6):1719–1724. doi: 10.1111/j.1471-4159.1980.tb11266.x. [DOI] [PubMed] [Google Scholar]
  21. Tamir H., Gershon M. D. Storage of serotonin and serotonin binding protein in synaptic vesicles. J Neurochem. 1979 Jul;33(1):35–44. doi: 10.1111/j.1471-4159.1979.tb11703.x. [DOI] [PubMed] [Google Scholar]
  22. Tamir H., Klein A., Rapport M. M. Serotonin binding protein:enhancement of binding by Fe2+ and inhibition of binding by drugs. J Neurochem. 1976 May;26(5):871–878. doi: 10.1111/j.1471-4159.1976.tb06467.x. [DOI] [PubMed] [Google Scholar]
  23. Tamir H., Liu K. P. On the nature of the interaction between serotonin and serotonin binding protein: effect of nucleotides, ions, and sulfhydryl reagents. J Neurochem. 1982 Jan;38(1):135–141. doi: 10.1111/j.1471-4159.1982.tb10864.x. [DOI] [PubMed] [Google Scholar]
  24. Theoharides T. C., Douglas W. W. Somatostatin induces histamine secretion from rat peritoneal mast cells. Endocrinology. 1978 May;102(5):1637–1640. doi: 10.1210/endo-102-5-1637. [DOI] [PubMed] [Google Scholar]
  25. Uvnäs B. Chemistry and storage function of mast cell granules. J Invest Dermatol. 1978 Jul;71(1):76–80. doi: 10.1111/1523-1747.ep12544425. [DOI] [PubMed] [Google Scholar]
  26. Yurt R., Austen K. F. Preparative purification of the rat mast cell chymase: characterization and interaction with granule components. J Exp Med. 1977 Nov 1;146(5):1405–1419. doi: 10.1084/jem.146.5.1405. [DOI] [PMC free article] [PubMed] [Google Scholar]

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