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. 1993 Apr 1;291(Pt 1):77–82. doi: 10.1042/bj2910077

Monoclonal antibodies to phosphatidylinositol 4-phosphate 5-kinase: distribution and intracellular localization of the C isoform.

C E Brooksbank 1, A Hutchings 1, G W Butcher 1, R F Irvine 1, N Divecha 1
PMCID: PMC1132483  PMID: 8385940

Abstract

We have raised a panel of monoclonal antibodies to PtdIns4P 5-kinase C purified from bovine brain [Divecha, Brooksbank and Irvine (1992) Biochem. J. 288, 637-642]. This panel includes antibodies which specifically recognize PtdIns4P 5-kinase C both in a native catalytically active condition, and/or when presented on Western blots. Some of the former antibodies will also inhibit PtdIns4P 5-kinase C activity. We have used the blotting antibodies to study the bovine tissue distribution of PtdIns4P 5-kinase C and its distribution in mammalian species. We have also studied its localization in Jurkat cells and found it to be predominantly bound to membranes, with only a minority localized to the cytoskeleton. Neither PtdIns4P 5-kinase activity nor PtdIns4P 5-kinase C, as detected by Western blotting, were increased in the cytoskeleton after stimulation of Jurkat cells with OKT3. These antibodies should prove to be extremely useful tools with which to study the regulation of PtdIns4P 5-kinase C.

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Selected References

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

  1. Alexander D. R., Hexham J. M., Lucas S. C., Graves J. D., Cantrell D. A., Crumpton M. J. A protein kinase C pseudosubstrate peptide inhibits phosphorylation of the CD3 antigen in streptolysin-O-permeabilized human T lymphocytes. Biochem J. 1989 Jun 15;260(3):893–901. doi: 10.1042/bj2600893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  3. Bazenet C. E., Ruano A. R., Brockman J. L., Anderson R. A. The human erythrocyte contains two forms of phosphatidylinositol-4-phosphate 5-kinase which are differentially active toward membranes. J Biol Chem. 1990 Oct 15;265(29):18012–18022. [PubMed] [Google Scholar]
  4. Berridge M. J., Irvine R. F. Inositol phosphates and cell signalling. Nature. 1989 Sep 21;341(6239):197–205. doi: 10.1038/341197a0. [DOI] [PubMed] [Google Scholar]
  5. Casadaban M. J., Cohen S. N. Lactose genes fused to exogenous promoters in one step using a Mu-lac bacteriophage: in vivo probe for transcriptional control sequences. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4530–4533. doi: 10.1073/pnas.76.9.4530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cochet C., Chambaz E. M. Catalytic properties of a purified phosphatidylinositol-4-phosphate kinase from rat brain. Biochem J. 1986 Jul 1;237(1):25–31. doi: 10.1042/bj2370025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cochet C., Filhol O., Payrastre B., Hunter T., Gill G. N. Interaction between the epidermal growth factor receptor and phosphoinositide kinases. J Biol Chem. 1991 Jan 5;266(1):637–644. [PubMed] [Google Scholar]
  8. Cockcroft S., Lamb J. R., Zanders E. D. Inositol lipid metabolism in human T lymphocytes activated via the T3 complex. Immunology. 1987 Feb;60(2):209–212. [PMC free article] [PubMed] [Google Scholar]
  9. Divecha N., Brooksbank C. E., Irvine R. F. Purification and characterization of phosphatidylinositol 4-phosphate 5-kinases. Biochem J. 1992 Dec 1;288(Pt 2):637–642. doi: 10.1042/bj2880637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Divecha N., Rhee S. G., Letcher A. J., Irvine R. F. Phosphoinositide signalling enzymes in rat liver nuclei: phosphoinositidase C isoform beta 1 is specifically, but not predominantly, located in the nucleus. Biochem J. 1993 Feb 1;289(Pt 3):617–620. doi: 10.1042/bj2890617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fraker P. J., Speck J. C., Jr Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphrenylglycoluril. Biochem Biophys Res Commun. 1978 Feb 28;80(4):849–857. doi: 10.1016/0006-291x(78)91322-0. [DOI] [PubMed] [Google Scholar]
  12. Galfrè G., Milstein C. Preparation of monoclonal antibodies: strategies and procedures. Methods Enzymol. 1981;73(Pt B):3–46. doi: 10.1016/0076-6879(81)73054-4. [DOI] [PubMed] [Google Scholar]
  13. Galfrè G., Milstein C., Wright B. Rat x rat hybrid myelomas and a monoclonal anti-Fd portion of mouse IgG. Nature. 1979 Jan 11;277(5692):131–133. doi: 10.1038/277131a0. [DOI] [PubMed] [Google Scholar]
  14. Gaudette D. C., Holub B. J. Effect of genistein, a tyrosine kinase inhibitor, on U46619-induced phosphoinositide phosphorylation in human platelets. Biochem Biophys Res Commun. 1990 Jul 16;170(1):238–242. doi: 10.1016/0006-291x(90)91265-t. [DOI] [PubMed] [Google Scholar]
  15. Gee N. S., Matsas R., Kenny A. J. A monoclonal antibody to kidney endopeptidase-24.11. Its application in immunoadsorbent purification of the enzyme and immunofluorescent microscopy of kidney and intestine. Biochem J. 1983 Aug 15;214(2):377–386. doi: 10.1042/bj2140377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Grondin P., Plantavid M., Sultan C., Breton M., Mauco G., Chap H. Interaction of pp60c-src, phospholipase C, inositol-lipid, and diacyglycerol kinases with the cytoskeletons of thrombin-stimulated platelets. J Biol Chem. 1991 Aug 25;266(24):15705–15709. [PubMed] [Google Scholar]
  17. Hawkins P. T., Jackson T. R., Stephens L. R. Platelet-derived growth factor stimulates synthesis of PtdIns(3,4,5)P3 by activating a PtdIns(4,5)P2 3-OH kinase. Nature. 1992 Jul 9;358(6382):157–159. doi: 10.1038/358157a0. [DOI] [PubMed] [Google Scholar]
  18. Husebye E. S., Flatmark T. Purification and kinetic properties of a soluble phosphatidylinositol-4-phosphate kinase of the bovine adrenal medulla with emphasis on its inhibition by calcium ions. Biochim Biophys Acta. 1989 Feb 9;1010(2):250–257. doi: 10.1016/0167-4889(89)90169-9. [DOI] [PubMed] [Google Scholar]
  19. Inokuchi S., Imboden J. B. Antigen receptor-mediated regulation of sustained polyphosphoinositide turnover in a human T cell line. Evidence for a receptor-regulated pathway for production of phosphatidylinositol 4,5-bisphosphate. J Biol Chem. 1990 Apr 15;265(11):5983–5989. [PubMed] [Google Scholar]
  20. Irvine R. F., Letcher A. J., Heslop J. P., Berridge M. J. The inositol tris/tetrakisphosphate pathway--demonstration of Ins(1,4,5)P3 3-kinase activity in animal tissues. Nature. 1986 Apr 17;320(6063):631–634. doi: 10.1038/320631a0. [DOI] [PubMed] [Google Scholar]
  21. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  22. Ling L. E., Schulz J. T., Cantley L. C. Characterization and purification of membrane-associated phosphatidylinositol-4-phosphate kinase from human red blood cells. J Biol Chem. 1989 Mar 25;264(9):5080–5088. [PubMed] [Google Scholar]
  23. Lundberg G. A., Jergil B., Sundler R. Phosphatidylinositol-4-phosphate kinase from rat brain. Activation by polyamines and inhibition by phosphatidylinositol 4,5-bisphosphate. Eur J Biochem. 1986 Dec 1;161(2):257–262. doi: 10.1111/j.1432-1033.1986.tb10441.x. [DOI] [PubMed] [Google Scholar]
  24. Moritz A., De Graan P. N., Ekhart P. F., Gispen W. H., Wirtz K. W. Purification of a phosphatidylinositol 4-phosphate kinase from bovine brain membranes. J Neurochem. 1990 Jan;54(1):351–354. doi: 10.1111/j.1471-4159.1990.tb13322.x. [DOI] [PubMed] [Google Scholar]
  25. Nahas N., Plantavid M., Mauco G., Chap H. Association of phosphatidylinositol kinase and phosphatidylinositol 4-phosphate kinase activities with the cytoskeleton in human platelets. FEBS Lett. 1989 Mar 27;246(1-2):30–34. doi: 10.1016/0014-5793(89)80247-9. [DOI] [PubMed] [Google Scholar]
  26. Nishizuka Y. The molecular heterogeneity of protein kinase C and its implications for cellular regulation. Nature. 1988 Aug 25;334(6184):661–665. doi: 10.1038/334661a0. [DOI] [PubMed] [Google Scholar]
  27. Payrastre B., Nievers M., Boonstra J., Breton M., Verkleij A. J., Van Bergen en Henegouwen P. M. A differential location of phosphoinositide kinases, diacylglycerol kinase, and phospholipase C in the nuclear matrix. J Biol Chem. 1992 Mar 15;267(8):5078–5084. [PubMed] [Google Scholar]
  28. Payrastre B., Plantavid M., Breton M., Chambaz E., Chap H. Relationship between phosphoinositide kinase activities and protein tyrosine phosphorylation in plasma membranes from A431 cells. Biochem J. 1990 Dec 15;272(3):665–670. doi: 10.1042/bj2720665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Payrastre B., van Bergen en Henegouwen P. M., Breton M., den Hartigh J. C., Plantavid M., Verkleij A. J., Boonstra J. Phosphoinositide kinase, diacylglycerol kinase, and phospholipase C activities associated to the cytoskeleton: effect of epidermal growth factor. J Cell Biol. 1991 Oct;115(1):121–128. doi: 10.1083/jcb.115.1.121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rhee S. G. Inositol phospholipids-specific phospholipase C: interaction of the gamma 1 isoform with tyrosine kinase. Trends Biochem Sci. 1991 Aug;16(8):297–301. doi: 10.1016/0968-0004(91)90122-c. [DOI] [PubMed] [Google Scholar]
  31. Rubin R., Hoek J. B. Ethanol-induced stimulation of phosphoinositide turnover and calcium influx in isolated hepatocytes. Biochem Pharmacol. 1988 Jun 15;37(12):2461–2466. doi: 10.1016/0006-2952(88)90374-7. [DOI] [PubMed] [Google Scholar]
  32. Smith C. D., Chang K. J. Regulation of brain phosphatidylinositol-4-phosphate kinase by GTP analogues. A potential role for guanine nucleotide regulatory proteins. J Biol Chem. 1989 Feb 25;264(6):3206–3210. [PubMed] [Google Scholar]
  33. 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]
  34. Thomas A. P., Marks J. S., Coll K. E., Williamson J. R. Quantitation and early kinetics of inositol lipid changes induced by vasopressin in isolated and cultured hepatocytes. J Biol Chem. 1983 May 10;258(9):5716–5725. [PubMed] [Google Scholar]
  35. Urumow T., Wieland O. H. A small G-protein involved in phosphatidylinositol-4-phosphate kinase activation. FEBS Lett. 1990 Apr 9;263(1):15–17. doi: 10.1016/0014-5793(90)80694-e. [DOI] [PubMed] [Google Scholar]
  36. Urumow T., Wieland O. H. Evidence for a cholera-toxin-sensitive G-protein involved in the regulation of phosphatidylinositol 4-phosphate kinase of rat liver membranes. Biochim Biophys Acta. 1988 Nov 18;972(2):232–238. doi: 10.1016/0167-4889(88)90121-8. [DOI] [PubMed] [Google Scholar]
  37. Urumow T., Wieland O. H. Purification and partial characterization of phosphatidylinositol-4-phosphate kinase from rat liver plasma membranes. Further evidence for a stimulatory G-protein. Biochim Biophys Acta. 1990 Apr 9;1052(1):152–158. doi: 10.1016/0167-4889(90)90070-t. [DOI] [PubMed] [Google Scholar]
  38. Van Dongen C. J., Zwiers H., Gispen W. H. Purification and partial characterization of the phosphatidylinositol 4-phosphate kinase from rat brain. Biochem J. 1984 Oct 1;223(1):197–203. doi: 10.1042/bj2230197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Van Rooijen L. A., Rossowska M., Bazan N. G. Inhibition of phosphatidylinositol-4-phosphate kinase by its product phosphatidylinositol-4,5-bisphosphate. Biochem Biophys Res Commun. 1985 Jan 16;126(1):150–155. doi: 10.1016/0006-291x(85)90584-4. [DOI] [PubMed] [Google Scholar]

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