Abstract
Phosphoinositide 3-kinase (PI3K) and its product phosphatidylinositol 3,4,5-trisphosphate (PIP3) play an essential role in the regulation of neutrophil functions by the chemoattractant formylmethionyl-leucylphenylalanine (FMLP). Here we show that permeabilization of human neutrophils leads to loss of cytosolic components, including PI3Kgamma, and causes the loss of FMLP-dependent production of PIP3. FMLP-sensitive synthesis of PIP3 could be restored by reconstitution of permeabilized neutrophils with recombinant PI3Kgamma. Admixture of recombinant phosphatidylinositol transfer protein (PITP) to the reconstitution cocktail produced a further increase of PIP3 synthesis, whereas pertussis toxin suppressed the FMLP-dependent production of PIP3. We conclude that FMLP-sensitive PIP3 formation in human neutrophils involves the FMLP receptor, heterotrimeric G-proteins of the Gi type, PI3Kgamma and PITP.
Full Text
The Full Text of this article is available as a PDF (190.1 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Arcaro A., Wymann M. P. Wortmannin is a potent phosphatidylinositol 3-kinase inhibitor: the role of phosphatidylinositol 3,4,5-trisphosphate in neutrophil responses. Biochem J. 1993 Dec 1;296(Pt 2):297–301. doi: 10.1042/bj2960297. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Avdi N. J., Winston B. W., Russel M., Young S. K., Johnson G. L., Worthen G. S. Activation of MEKK by formyl-methionyl-leucyl-phenylalanine in human neutrophils. Mapping pathways for mitogen-activated protein kinase activation. J Biol Chem. 1996 Dec 27;271(52):33598–33606. doi: 10.1074/jbc.271.52.33598. [DOI] [PubMed] [Google Scholar]
- Baggiolini M., Dewald B., Schnyder J., Ruch W., Cooper P. H., Payne T. G. Inhibition of the phagocytosis-induced respiratory burst by the fungal metabolite wortmannin and some analogues. Exp Cell Res. 1987 Apr;169(2):408–418. doi: 10.1016/0014-4827(87)90201-1. [DOI] [PubMed] [Google Scholar]
- Cunningham E., Tan S. K., Swigart P., Hsuan J., Bankaitis V., Cockcroft S. The yeast and mammalian isoforms of phosphatidylinositol transfer protein can all restore phospholipase C-mediated inositol lipid signaling in cytosol-depleted RBL-2H3 and HL-60 cells. Proc Natl Acad Sci U S A. 1996 Jun 25;93(13):6589–6593. doi: 10.1073/pnas.93.13.6589. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cunningham E., Thomas G. M., Ball A., Hiles I., Cockcroft S. Phosphatidylinositol transfer protein dictates the rate of inositol trisphosphate production by promoting the synthesis of PIP2. Curr Biol. 1995 Jul 1;5(7):775–783. doi: 10.1016/s0960-9822(95)00154-0. [DOI] [PubMed] [Google Scholar]
- Fensome A., Cunningham E., Prosser S., Tan S. K., Swigart P., Thomas G., Hsuan J., Cockcroft S. ARF and PITP restore GTP gamma S-stimulated protein secretion from cytosol-depleted HL60 cells by promoting PIP2 synthesis. Curr Biol. 1996 Jun 1;6(6):730–738. doi: 10.1016/s0960-9822(09)00454-0. [DOI] [PubMed] [Google Scholar]
- Hay J. C., Fisette P. L., Jenkins G. H., Fukami K., Takenawa T., Anderson R. A., Martin T. F. ATP-dependent inositide phosphorylation required for Ca(2+)-activated secretion. Nature. 1995 Mar 9;374(6518):173–177. doi: 10.1038/374173a0. [DOI] [PubMed] [Google Scholar]
- Hay J. C., Martin T. F. Phosphatidylinositol transfer protein required for ATP-dependent priming of Ca(2+)-activated secretion. Nature. 1993 Dec 9;366(6455):572–575. doi: 10.1038/366572a0. [DOI] [PubMed] [Google Scholar]
- Kauffmann-Zeh A., Thomas G. M., Ball A., Prosser S., Cunningham E., Cockcroft S., Hsuan J. J. Requirement for phosphatidylinositol transfer protein in epidermal growth factor signaling. Science. 1995 May 26;268(5214):1188–1190. doi: 10.1126/science.7761838. [DOI] [PubMed] [Google Scholar]
- Lopez-Ilasaca M., Crespo P., Pellici P. G., Gutkind J. S., Wetzker R. Linkage of G protein-coupled receptors to the MAPK signaling pathway through PI 3-kinase gamma. Science. 1997 Jan 17;275(5298):394–397. doi: 10.1126/science.275.5298.394. [DOI] [PubMed] [Google Scholar]
- Martin T. Intracellular signalling. New directions for phosphatidylinositol transfer. Curr Biol. 1995 Sep 1;5(9):990–992. doi: 10.1016/s0960-9822(95)00196-5. [DOI] [PubMed] [Google Scholar]
- Ohashi M., Jan de Vries K., Frank R., Snoek G., Bankaitis V., Wirtz K., Huttner W. B. A role for phosphatidylinositol transfer protein in secretory vesicle formation. Nature. 1995 Oct 12;377(6549):544–547. doi: 10.1038/377544a0. [DOI] [PubMed] [Google Scholar]
- Premack B. A., Schall T. J. Chemokine receptors: gateways to inflammation and infection. Nat Med. 1996 Nov;2(11):1174–1178. doi: 10.1038/nm1196-1174. [DOI] [PubMed] [Google Scholar]
- Stephens L. R., Hughes K. T., Irvine R. F. Pathway of phosphatidylinositol(3,4,5)-trisphosphate synthesis in activated neutrophils. Nature. 1991 May 2;351(6321):33–39. doi: 10.1038/351033a0. [DOI] [PubMed] [Google Scholar]
- Stephens L., Eguinoa A., Corey S., Jackson T., Hawkins P. T. Receptor stimulated accumulation of phosphatidylinositol (3,4,5)-trisphosphate by G-protein mediated pathways in human myeloid derived cells. EMBO J. 1993 Jun;12(6):2265–2273. doi: 10.1002/j.1460-2075.1993.tb05880.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stephens L., Jackson T., Hawkins P. T. Synthesis of phosphatidylinositol 3,4,5-trisphosphate in permeabilized neutrophils regulated by receptors and G-proteins. J Biol Chem. 1993 Aug 15;268(23):17162–17172. [PubMed] [Google Scholar]
- Stoyanov B., Volinia S., Hanck T., Rubio I., Loubtchenkov M., Malek D., Stoyanova S., Vanhaesebroeck B., Dhand R., Nürnberg B. Cloning and characterization of a G protein-activated human phosphoinositide-3 kinase. Science. 1995 Aug 4;269(5224):690–693. doi: 10.1126/science.7624799. [DOI] [PubMed] [Google Scholar]
- Thelen M., Wymann M. P., Langen H. Wortmannin binds specifically to 1-phosphatidylinositol 3-kinase while inhibiting guanine nucleotide-binding protein-coupled receptor signaling in neutrophil leukocytes. Proc Natl Acad Sci U S A. 1994 May 24;91(11):4960–4964. doi: 10.1073/pnas.91.11.4960. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thomas G. M., Cunningham E., Fensome A., Ball A., Totty N. F., Truong O., Hsuan J. J., Cockcroft S. An essential role for phosphatidylinositol transfer protein in phospholipase C-mediated inositol lipid signaling. Cell. 1993 Sep 10;74(5):919–928. doi: 10.1016/0092-8674(93)90471-2. [DOI] [PubMed] [Google Scholar]
- Wirtz K. W. Phospholipid transfer proteins. Annu Rev Biochem. 1991;60:73–99. doi: 10.1146/annurev.bi.60.070191.000445. [DOI] [PubMed] [Google Scholar]