Isolation and Regulation of the cGMP-lnhibited cAMP Phosphodiesterase in Human Erythroleukemia Cells

S B Sheth; K J Brennan; R Biradavolu; R W Colman

doi:10.1055/s-0038-1655924

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 1997; 77(01): 155-162
DOI: 10.1055/s-0038-1655924

Platelets

Schattauer GmbH Stuttgart

Isolation and Regulation of the cGMP-lnhibited cAMP Phosphodiesterase in Human Erythroleukemia Cells

Autoren

S B Sheth

The Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA, USA
K J Brennan

The Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA, USA
R Biradavolu

The Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA, USA
R W Colman

The Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA, USA

Abstract

als PDF herunterladen

Summary

The predominant cAMP phosphodiesterase in human platelets is the low K_m cGMP-inhibited phosphodiesterase (PDE 3A). We have isolated native PDE3A from platelets and human erythroleukemia (HEL) cells and studied its kinetics. The platelet and HEL cell enzymes hydrolyze cAMP with K_m = 0.5 μM. Incubation of cell supernatant with cAMP dependent protein kinase resulted in a rapid increase in activity within minutes, which resulted from a 2-fold decrease in K_m with no increase in V_max. HEL cells grown for 24 h in the presence of 50 (iM forskolin, an adenylate cyclase activator, demonstrate further increase in PDE3A of 274% of control (p = 0.03). Cells incubated with forskolin and cycloheximide or actinomycin D demonstrated no increase suggesting that cAMP stimulates PDE3A synthesis by transcriptional regulation. The results indicate that cAMP affects both the short and long-term regulation of PDE3A. The latter effect may play a role in the developing hematopoietic cell and the cardiovascular system to regulate cAMP levels.

PDF (2734 kb)

Referenzen

References
1 Hidaka H, Endo T. Human blood platelet 3’5’-cyclic nucleotide phosphodiesterase. Isolation of low-km and high-km phosphodiesterase. Biochim Biophys Acta 1976; 429: 485-497
2 Sheth SB, Colman RW. Regulatory and catalytic domains of platelet cAMP phosphodiesterases: targets for drug design. [Review]. Semin Heme 1996; 110-119
3 Grant PG, Colman RW. Purification and characterization of a human platelet cyclic nucleotide phosphodiesterase. Biochem 1984; 23: 1801-1807
4 Honerjager P. Pharmacology of positive inotropic phosphodiesterase III inhibitors. Eur Heart J 1989; 10 Suppl C 25-31
5 Weishaar RE, Kobylarz-Singer DC, Kaplan HR. Subclasses of cyclic AMP phosphodiesterase in cardiac muscle. J Mol Cell Card 1987; 19: 1025-1036
6 Weishaar RE, Kobylarz-Singer DC, Quade MM, Steffen RP, Kaplan HR. Multiple molecular forms of phosphodiesterase and the regulation of cardiac muscle contractility. J Cyc Nuc Prot Phos Res 1986; 11: 513-527
7 Weishaar RE, Burrows SD, Kobylarz DC, Quade MM, Evans DB. Multiple molecular forms of cyclic nucleotide phosphodiesterase in cardiac and smooth muscle and in platelets. Isolation, characterization, and effects of various reference phosphodiesterase inhibitors and cardiotonic agents. Biochem Pharm 1986; 35: 787-800
8 Suttorp N, Weber U, Welsch T, Schudt C. Role of phosphodiesterases in the regulation of endothelial permeability in vitro. J Clin Invest 1993; 91: 1421-1428
9 Degerman E, Moos jr M, Rascón A, Vasta V, Meacci E, Smith CJ, Andersson KE, lingersn S, Belfrage P, Manganiello V. Single-step affinity p urification partial structure and properties of human platelet cGMP inhibited cAMP phosphodiesterase. Biochim Biophys Acta Protein Struct Mol Enzymol 1994; 1205: 189-198
10 Charbonneau H. Structure-function relationships among cyclic nucleotide phophodiesterases. In: Cyclic Nucleotide Phosphodiesterases: Structure, Regulation and Drug Action. Beavo J, Houslay MD. (eds) New York: John Wiley and Sons; 1990. pp 267-296
11 Tabilio A, Rosa JP, Testa U, Kieffer N, Nurden AT, Del Canizo MC, Breton-Gorius J, Vainchender W. Expression of platelet membrane glycoproteins and α-granule proteins by a human erythroleukemia cell line (HEL). EMBOJ 1984; 03: 453-459
12 Ashby B, Almonor GO, Wernick E, Selak MA. Prostaglandin-concentration-dependent desensitization of adenylate cyclase in human erythroleu-kaemia (HEL)cells is abolished by pertussis toxin and enhanced by induction by dimethyl sulphoxide. Biochem J 1991; 280: 801-804
13 Macphee CH, Reifsnyder DH, Moore TA, Beavo JA. Intact cell and cell-free phosphorylation and concomitant activation of a low Km cAMP phosphodiesterase found in human platelets. J Cyc Nuc Prot Phos Res 1986; 11: 487-496
14 Grant PG, Mannarino AF, Colman RW. cAMP-mediated phosphorylation of the low-Km cAMP phosphodiesterase markedly stimulates its catalytic activity. Proc Natl Acad Sci USA 1988; 85: 9071-9075
15 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685
16 Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 1979; 76: 4350-4354
17 Torphy TJ, Zhou HL, Foley JJ, Sarau HM, Manning CD, Barnette MS. Salbutamol up-regulates PDE4 activity and induces a heterologous desensitization of U937 cells to prostaglandin E₂ . Implications for the therapeutic use of beta-adrenoceptor agonists. J Biol Chem 1996; 270: 23598-23604
18 Sinha AK, Shattil SJ, Colman RW. Cyclic AMP metabolism in cholesterol-rich platelets. J Biol Chem 1977; 252: 3310-3314
19 Torphy TJ, Cieslinski LB. Characterization and selective inhibition of cyclic nucleotide phosphodiesterase isozymes in canine tracheal smooth muscle. Mol Pharmacol 1990; 37: 206-214
20 Thompson WJ, Terasaki WL, Epstein PM, Strada SJ. Assay of cyclic nucleotide phosphodiesterase and resolution of multiple molecular forms of the enzyme [Review]. Adv Cyc Nuc Res 1979; 10: 69-92
21 Macphee CH, Reifsnyder DH, Moore TA, Lerea KM, Beavo JA. Phosphorylation results in activation of a cAMP phosphodiesterase in human platelets. J Biol Chem 1988; 263: 10353-10358
22 Macphee CH, Harrison SA, Beavo JA. Immunological identification of the major platelet low-Km cAMP phosphodiesterase: probable target for antithrombotic agents. Proc Natl Acad Sci USA 1986; 83: 6660-6663
23 Rascon A, Degerman E, Taira M, Meacci E, Smith CJ, Manganiello V, Belfrage P, Tornqvist H. Identification of the phosphorylation site in vitro for cAMP-dependent protein kinase on the rat adipocyte cGMP-inhibited cAMP phosphodiesterase. J Biol Chem 1994; 269: 11962-11966
24 Rahn T, Ronnstrand L, Leroy MJ, Wemsted C, Tornqvist H, Manganiello VC, Belfrage P, Degerman E. Identification of the site in the cGMP-inhibited phosphodiesterase phosphorylated in adipocytes in response to insulin and isoproterenol. J Biol Chem 1996; 271: 11575-11580
25 Sette C, Vicini E, Conti M. The ratPDE3/IVd phosphodiesterase gene codes for multiple proteins differentially activated by cAMP-dependent protein kinase [published erratum appears in J Biol Chem 1994 Aug 12; 269(32): 20806]. J Biol Chem 1994; 269: 18271-18274
26 Gettys TW, Blackmore PF, Redmon JB, Beebe SJ, Corbin JD. Short-term feedback regulation of cAMP by accelerated degradation in rat tissues. J Biol Chem 1987; 262: 333-339
27 Cheung P, Xu H, McLaughlin MM, Ghazeleh FA, Livi GP, Colman RW. Human platelet cGI-PDE: Expression in yeast and localization of the catalytic domain by deletion mutagenesis. Blood 1996; 88: 1321-1329
28 Meacci E, Taira M, Moos jr M, Smith CJ, Movsesian MA, Degerman E, Belfrage P, Manganiello V. Molecular cloning and expression of human myocardial cGMP-inhibited cAMP phosphodiesterase. Proc Natl Acad Sci USA 1992; 89: 3721-3725
29 Taira M, Hockman SC, Calvo JC, Belfrage P, Manganiello VC. Molecular cloning of the rat adipocyte hormone-sensitive cyclic GMP-inhibited cyclic nucleotide phosphodiesterase. J Biol Chem 1993; 268: 18573-18579
30 Mochly-Rosen D. Localization of protein kinases by anchoring proteins: a theme in signal transduction. [Review]. Science 1995; 268: 247-251
31 Richardson MD, Goka TJ, Barber R, Butcher RW. Growth of S49 wild type cells in 3 nM epinephrine increases cyclic AMP phosphodiesterase activity. Life Sciences 1994; 54: 863-875
32 Torphy TJ, Zhou HL, Cieslinski LB. Stimulation of beta adrenoceptors in a human monocyte cell line (U937) up-regulates cyclic AMP-specific phosphodiesterase activity. J Pharmacol Exp Ther 1992; 263: 1195-1205
33 Mazur EM, Rosmarin AG, Sohl PA, Newton JL, Narendran A. Analysis of the mechanism of anagrelide-induced thrombocytopenia in humans. Blood 1992; 79: 1931-1937