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. 1991 Apr 1;275(Pt 1):127–131. doi: 10.1042/bj2750127

Relationship between arachidonate release and exocytosis in permeabilized human neutrophils stimulated with formylmethionyl-leucyl-phenylalanine (fMetLeuPhe), guanosine 5'-[gamma-thio]triphosphate (GTP[S]) and Ca2+.

S Cockcroft 1
PMCID: PMC1150022  PMID: 1902082

Abstract

The role of two G-proteins, Gp and Ge, in the stimulus-secretion pathway has been proposed on the basis of studies where GTP analogues have been introduced into permeabilized cell preparations. In this study, evidence is provided that two G-proteins are also involved when a receptor-directed agonist is used. Intact human neutrophils were made refractory to formylmethionyl-leucyl-phenylalanine (fMetLeuPhe) stimulation by metabolic inhibition and then permeabilized with streptolysin O to compare the intracellular requirements for exocytosis from specific and azurophilic granules and arachidonate release. In the presence of 1 microM-Ca2+ and 1 mM-MgATP, fMetLeuPhe or guanosine 5'-[gamma-thio]triphosphate (GTP[S]) induce secretion from both granule types as well as arachidonate release. Secretion and arachidonate release owing to fMetLeuPhe can occur in the absence of ATP, conditions under which G-protein-mediated activation of phospholipase C is suppressed. GTP[S]-induced secretion can also occur in the absence of MgATP, but GTP[S]-induced arachidonate release cannot. It is concluded that fMetLeuPhe, like GTP[S], stimulates secretion by interacting with another G-protein-mediated reaction apart from Gp. Evidence is provided that a possible target for the second G-protein-mediated reaction involved in fMetLeuPhe-induced secretion (but not GTP[S]-induced secretion) is phospholipase A2.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barrowman M. M., Cockcroft S., Gomperts B. D. Differential control of azurophilic and specific granule exocytosis in Sendai-virus-permeabilized rabbit neutrophils. J Physiol. 1987 Feb;383:115–124. doi: 10.1113/jphysiol.1987.sp016399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barrowman M. M., Cockcroft S., Gomperts B. D. Two roles for guanine nucleotides in the stimulus-secretion sequence of neutrophils. Nature. 1986 Feb 6;319(6053):504–507. doi: 10.1038/319504a0. [DOI] [PubMed] [Google Scholar]
  3. Bennett J. P., Cockcroft S., Gomperts B. D. Use of cytochalasin B to distinguish between early and late events in neutrophil activation. Biochim Biophys Acta. 1980 Oct 2;601(3):584–591. doi: 10.1016/0005-2736(80)90560-x. [DOI] [PubMed] [Google Scholar]
  4. Bittner M. A., Holz R. W., Neubig R. R. Guanine nucleotide effects on catecholamine secretion from digitonin-permeabilized adrenal chromaffin cells. J Biol Chem. 1986 Aug 5;261(22):10182–10188. [PubMed] [Google Scholar]
  5. Bokoch G. M., Gilman A. G. Inhibition of receptor-mediated release of arachidonic acid by pertussis toxin. Cell. 1984 Dec;39(2 Pt 1):301–308. doi: 10.1016/0092-8674(84)90008-4. [DOI] [PubMed] [Google Scholar]
  6. Burch R. M., Axelrod J. Dissociation of bradykinin-induced prostaglandin formation from phosphatidylinositol turnover in Swiss 3T3 fibroblasts: evidence for G protein regulation of phospholipase A2. Proc Natl Acad Sci U S A. 1987 Sep;84(18):6374–6378. doi: 10.1073/pnas.84.18.6374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Burch R. M., Luini A., Axelrod J. Phospholipase A2 and phospholipase C are activated by distinct GTP-binding proteins in response to alpha 1-adrenergic stimulation in FRTL5 thyroid cells. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7201–7205. doi: 10.1073/pnas.83.19.7201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Churcher Y., Allan D., Gomperts B. D. Relationship between arachidonate generation and exocytosis in permeabilized mast cells. Biochem J. 1990 Feb 15;266(1):157–163. doi: 10.1042/bj2660157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Churcher Y., Kramer K. M., Gomperts B. D. Evidence for protein dephosphorylation as a permissive step in GTP-gamma-S-induced exocytosis from permeabilized mast cells. Cell Regul. 1990 Jun;1(7):523–530. doi: 10.1091/mbc.1.7.523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cockcroft S. Ca2+-dependent conversion of phosphatidylinositol to phosphatidate in neutrophils stimulated with fMet-Leu-Phe or ionophore A23187. Biochim Biophys Acta. 1984 Aug 15;795(1):37–46. doi: 10.1016/0005-2760(84)90102-4. [DOI] [PubMed] [Google Scholar]
  11. Cockcroft S., Howell T. W., Gomperts B. D. Two G-proteins act in series to control stimulus-secretion coupling in mast cells: use of neomycin to distinguish between G-proteins controlling polyphosphoinositide phosphodiesterase and exocytosis. J Cell Biol. 1987 Dec;105(6 Pt 1):2745–2750. doi: 10.1083/jcb.105.6.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cockcroft S., Stutchfield J. ATP stimulates secretion in human neutrophils and HL60 cells via a pertussis toxin-sensitive guanine nucleotide-binding protein coupled to phospholipase C. FEBS Lett. 1989 Mar 13;245(1-2):25–29. doi: 10.1016/0014-5793(89)80184-x. [DOI] [PubMed] [Google Scholar]
  13. Cockcroft S., Stutchfield J. The receptors for ATP and fMetLeuPhe are independently coupled to phospholipases C and A2 via G-protein(s). Relationship between phospholipase C and A2 activation and exocytosis in HL60 cells and human neutrophils. Biochem J. 1989 Nov 1;263(3):715–723. doi: 10.1042/bj2630715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Eberhard D. A., Cooper C. L., Low M. G., Holz R. W. Evidence that the inositol phospholipids are necessary for exocytosis. Loss of inositol phospholipids and inhibition of secretion in permeabilized cells caused by a bacterial phospholipase C and removal of ATP. Biochem J. 1990 May 15;268(1):15–25. doi: 10.1042/bj2680015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Falloon J., Malech H., Milligan G., Unson C., Kahn R., Goldsmith P., Spiegel A. Detection of the major pertussis toxin substrate of human leukocytes with antisera raised against synthetic peptides. FEBS Lett. 1986 Dec 15;209(2):352–356. doi: 10.1016/0014-5793(86)81141-3. [DOI] [PubMed] [Google Scholar]
  16. Haslam R. J., Davidson M. M. Guanine nucleotides decrease the free [Ca2+] required for secretion of serotonin from permeabilized blood platelets. Evidence of a role for a GTP-binding protein in platelet activation. FEBS Lett. 1984 Aug 20;174(1):90–95. doi: 10.1016/0014-5793(84)81084-4. [DOI] [PubMed] [Google Scholar]
  17. Howell T. W., Cockcroft S., Gomperts B. D. Essential synergy between Ca2+ and guanine nucleotides in exocytotic secretion from permeabilized rat mast cells. J Cell Biol. 1987 Jul;105(1):191–197. doi: 10.1083/jcb.105.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Howell T. W., Kramer I. M., Gomperts B. D. Protein phosphorylation and the dependence on Ca2+ and GTP-gamma-S for exocytosis from permeabilised mast cells. Cell Signal. 1989;1(2):157–163. doi: 10.1016/0898-6568(89)90005-3. [DOI] [PubMed] [Google Scholar]
  19. Jelsema C. L., Axelrod J. Stimulation of phospholipase A2 activity in bovine rod outer segments by the beta gamma subunits of transducin and its inhibition by the alpha subunit. Proc Natl Acad Sci U S A. 1987 Jun;84(11):3623–3627. doi: 10.1073/pnas.84.11.3623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Knight D. E., Scrutton M. C. Effects of guanine nucleotides on the properties of 5-hydroxytryptamine secretion from electropermeabilised human platelets. Eur J Biochem. 1986 Oct 1;160(1):183–190. doi: 10.1111/j.1432-1033.1986.tb09956.x. [DOI] [PubMed] [Google Scholar]
  21. Knight D. E., Scrutton M. C. Gaining access to the cytosol: the technique and some applications of electropermeabilization. Biochem J. 1986 Mar 15;234(3):497–506. doi: 10.1042/bj2340497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Morgan A., Burgoyne R. D. Stimulation of Ca2(+)-independent catecholamine secretion from digitonin-permeabilized bovine adrenal chromaffin cells by guanine nucleotide analogues. Relationship to arachidonate release. Biochem J. 1990 Jul 15;269(2):521–526. doi: 10.1042/bj2690521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Murphy P. M., Eide B., Goldsmith P., Brann M., Gierschik P., Spiegel A., Malech H. L. Detection of multiple forms of Gi alpha in HL60 cells. FEBS Lett. 1987 Aug 31;221(1):81–86. doi: 10.1016/0014-5793(87)80356-3. [DOI] [PubMed] [Google Scholar]
  24. Nakashima S., Nagata K., Ueeda K., Nozawa Y. Stimulation of arachidonic acid release by guanine nucleotide in saponin-permeabilized neutrophils: evidence for involvement of GTP-binding protein in phospholipase A2 activation. Arch Biochem Biophys. 1988 Mar;261(2):375–383. doi: 10.1016/0003-9861(88)90353-0. [DOI] [PubMed] [Google Scholar]
  25. Narasimhan V., Holowka D., Baird B. A guanine nucleotide-binding protein participates in IgE receptor-mediated activation of endogenous and reconstituted phospholipase A2 in a permeabilized cell system. J Biol Chem. 1990 Jan 25;265(3):1459–1464. [PubMed] [Google Scholar]
  26. Okano Y., Yamada K., Yano K., Nozawa Y. Guanosine 5'-(gamma-thio)triphosphate stimulates arachidonic acid liberation in permeabilized rat peritoneal mast cells. Biochem Biophys Res Commun. 1987 Jun 30;145(3):1267–1275. doi: 10.1016/0006-291x(87)91574-9. [DOI] [PubMed] [Google Scholar]
  27. Peppers S. C., Holz R. W. Catecholamine secretion from digitonin-treated PC12 cells. Effects of Ca2+, ATP, and protein kinase C activators. J Biol Chem. 1986 Nov 5;261(31):14665–14669. [PubMed] [Google Scholar]
  28. Segal A. W., Dorling J., Coade S. Kinetics of fusion of the cytoplasmic granules with phagocytic vacuoles in human polymorphonuclear leukocytes. Biochemical and morphological studies. J Cell Biol. 1980 Apr;85(1):42–59. doi: 10.1083/jcb.85.1.42. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Smolen J. E., Stoehr S. J., Boxer L. A. Human neutrophils permeabilized with digitonin respond with lysosomal enzyme release when exposed to micromolar levels of free calcium. Biochim Biophys Acta. 1986 Apr 8;886(1):1–17. doi: 10.1016/0167-4889(86)90205-3. [DOI] [PubMed] [Google Scholar]
  30. Smolen J. E., Stoehr S. J. Guanine nucleotides reduce the free calcium requirement for secretion of granule constituents from permeabilized human neutrophils. Biochim Biophys Acta. 1986 Nov 28;889(2):171–178. doi: 10.1016/0167-4889(86)90101-1. [DOI] [PubMed] [Google Scholar]
  31. Stutchfield J., Cockcroft S. Guanine nucleotides stimulate polyphosphoinositide phosphodiesterase and exocytotic secretion from HL60 cells permeabilized with streptolysin O. Biochem J. 1988 Mar 1;250(2):375–382. doi: 10.1042/bj2500375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Vallar L., Biden T. J., Wollheim C. B. Guanine nucleotides induce Ca2+-independent insulin secretion from permeabilized RINm5F cells. J Biol Chem. 1987 Apr 15;262(11):5049–5056. [PubMed] [Google Scholar]
  33. Volterra A., Siegelbaum S. A. Role of two different guanine nucleotide-binding proteins in the antagonistic modulation of the S-type K+ channel by cAMP and arachidonic acid metabolites in Aplysia sensory neurons. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7810–7814. doi: 10.1073/pnas.85.20.7810. [DOI] [PMC free article] [PubMed] [Google Scholar]

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