Degradation of extracellular nucleotides and their analogs in HeLa and HUVEC cell cultures

• Autorzy: Gendaszewska-Darmach E. , Maszewska M. , Zaklos M. , Koziolkiewicz M.
• Języki publikacji: EN

Abstrakty

EN

The use of nucleotides and their analogs in the pharmacological studies of nucleo­tide receptors (P2 class) should be preceded by detailed studies on their degradation connected with ecto-enzymes of a given cell type. In the present studies we have ana­lyzed stability of some phosphorothioate and phosphonate analogs of ATP and ADP in the HeLa epitheloid carcinoma and endothelial HUVEC cells cultures. Our studies have revealed that ecto-nucleotide pyrophosphatase (E-NPP) is one of the main en­zymes involved in the extracellular degradation of ATP and other nucleotides in the HeLa cells. On the other hand, the ecto-ATPDase is responsible for the hydrolysis of extracellular nucleotides in human endothelial cell cultures, while the E-NPP-like en­zymes of the HUVEC cells are not essential to this degradation. The concerted action of the aforementioned ecto-enzymes and nucleotide pyrophosphatase, 5 -nucleo- tidase and adenosine deaminase present in fetal bovine serum (FBS) supplied to the culture medium, results in partial or complete degradation of the phosphorothioate (ATPγS) and phosphonate analogs of adenosine nucleotides (α,β-methylene-ATP and β,γ-methylene-ATP) in the cell cultures. Only ADPβS appears to be resistant to these enzymes. The influence of some nucleotides and their analogs on the proliferation of the HeLa cells in presence or absence of FBS is also discussed.

Słowa kluczowe

EN

nucleotide analogue; HeLa cell; ectoenzyme; degradation; extracellular nucleotide; nucleotide pyrophosphatase

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2003
• Tom: 50
• Numer: 4
• Opis fizyczny: p.973-984,fig.,ref.

Twórcy

autor

Gendaszewska-Darmach E.

• Polish Academy of Sciences, Lodz, Poland

autor

Maszewska M.

autor

Zaklos M.

autor

Koziolkiewicz M.

Bibliografia

• Birk AV, Bubman D, Broekman MJ, Robertson HD, Drosopoulos JH, Marcus AJ, Szeto HH. (2002) Role of a novel soluble nucleotide phosphohydrolase from sheep plasma in inhibition of platelet reactivity: hemostasis, thrombosis and vascular biology. J Lab Clin Med.; 139: 116-24.
• Burnstock G. (2002) Purinergic signaling and vascular cell proliferation and death. Arterioscler Thromb Vasc Biol. ; 22: 364-73.
• Claes P, Grobben B, van Kolen K, Roymans D, Slegers H. (2001) P2YAC- receptor agonists enhance the proliferation of rat C6 glioma cells through activation of the p42/44 mitogen-activated protein kinase. Br J Pharmacol.; 134: 402-8.
• Czajkowski R, Baranska J. (2002) Cross-talk between the ATP and ADP nucleotide receptor signalling pathways in glioma C6 cells. Acta Biochim Polon.; 49: 877-89.
• Frittitta L, Camastra S, Baratta R, Constanzo BV, D'Adamo M, Graci S, Spampinato D, Maddux BA, Vigneri R, Ferrannini E, Trischitta V. (1999) A soluble PC-1 circulates in human plasma: relationship with insulin resistance and associated abnormalities. J Clin EndocrinolMetab.; 84: 3620-5.
• Gijsbers R, Ceulemans H, Stalmans W, Bollen M. (2001) Structural and catalytic similarities between nucleotide pyrophosphatase/phosphodiesterase and alkaline phosphatase. J Biol Chem.; 276: 1361-8.
• Goding JW, Terkeltaub R, Maurice M, Deterre P, Sali A, Belli S. (1998) Ecto-phosphodiesterase/pyrophosphatase of lymphocytes and non-lymphoid cells: structure and function of the PC-1 family. Immunol Rev.; 161: 11-26.
• Goding JW, Grobben B, Slegers H. (2003) Physiological and pathophysiological functions of the ecto-nucleotide pyrophosphatase/phosphodiesterase family. Biochim Biophys Acta.; 1638: 1-19.
• Grobben B, Anciaux K, Roymans D, Stefan C, Bollen M, Esmans EL, Slegers H. (1999) An ecto-nucleotide pyrophosphatase is one of the main enzymes involved in the extracellular metabolism of ATP in rat C6 glioma. J Neurochem.; 72: 826-34.
• Hansen MB, Nielsen SE, Berg K. (1989) Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J Immunol Methods.; 119: 203-10.
• Hou M, Harden K, Kuhn CM, Baldetorp B, Lazarowski E, Pendergast W, Moller S, Edvinsson L, Erlinge D. (2002) UDP acts as a growth factor for vascular smooth muscle cells by activation of P2Y6 receptors. Am J Physiol Heart Circ Physiol.; 282: H784-92.
• Koziolkiewicz M, Gendaszewska E, Maszewska M, Stein CA, Stec WJ. (2001) The mononucleotide-dependent, nonantisense mechanism of action of phosphodiester and phosphorothioate oligonucleotides depends upon the activity of an ecto-5'-nucleotidase. Blood.; 98: 995-1002.
• Lazarowski ER, Homolya L, Boucher RC, Harden TK. (1997) Identification of an ecto-nucleoside diphosphokinase and its contribution to interconversion of P2 receptor agonists. J Biol Chem.; 272: 20402-7.
• Lazarowski ER, Boucher RC, Harden TK. (2000) Constitutive release of ATP and evidence for major contribution of ecto- nucleotide pyrophosphatase and nucleoside diphosphokinase to extracellular nucleotide concentration. J Biol Chem. ; 275: 31061-8.
• Mateo J, Rotllan P, Miras-Portugal MT. (1996) Suramin - a powerful inhibitor of neural ectoadenosine polyphosphate hydrolase. Br J Pharmacol.; 119: 1-2.
• Nobes CD, Hall A. (1999) Rho GTPases control polarity, protrusion and adhesion during cell movement. J Cell Biol.; 144: 1235-44.
• Picher M, Sevigny J, D'Orleans-Juste P, Beaudoin AR. (1996) Hydrolysis of P2-purinoceptor agonists by a purified ectonucleotidase from the bovine aorta, the ATP-diphosphohydrolase. Biochem Pharmacol.; 51: 1453-60.
• Ralevic V, Burnstock G. (1998) Receptors for purines and pyrimidines. Pharmacol Rev.; 50: 413-92.
• Stefan C, Gijsbers R, Stalmans W, Bollen M. (1999) Differential regulation of the expression of nucleotide pyrophosphatase and phosphodiesterase in rat liver. Biochim Biophys Acta.; 1450: 45-52.
• Stracke ML, Krutzsch HC, Unsworth EJ, Arestad A, Cioce V, Schiffmann E, Liotta LA. (1992) Identification, purification, and partial sequence analysis of autotaxin, a novel motility-stimulating protein. J Biol Chem.; 267: 2524-9.
• Tokumura A, Majima E, Kariya Y, Tominaga K, Kogure K, Yasuda K, Fukuzawa K. (2002) Identification of human plasma lysophospholipase D, a lysophosphatidic acid-producing enzyme as autotaxin, a multi-functional phosphodiesterase. J Biol Chem.; 277: 39436-42.
• Umezu-Goto M, Kishi Y, Taira A, Hama K, Dohmae N, Takio K, Yamori T, Mills GB, Inoue K, Aoki J, Arai H. (2002) Autotaxin has lysophospholipase D activity leading to tumor cell growth and motility by lysophosphatidic acid production. J Cell Biol.; 158: 227-33.
• Weisman GA, Lustig KD, Lane E, Huamg NN, Belzer I, Friedberg I. (1988) Growth inhibition of transformed mouse fibroblasts by adenine nucleotides occurs via generation of extracellular adenosine. J Biol Chem.; 263: 12367-72.
• Williams M, Jarvis MF. (2000) Purinergic and pyrimidinergic receptors as potential drug targets. Biochem Pharmacol.; 59: 1173-85.
• Yegutkin GG, Henttinen T, Samburski SS, Spychala J, Jalkanen S. (2002) The evidence for two opposite, ATP-generating and ATP-consuming extracellular pathways on endothelial and lymphoid cells. Biochem J.; 367: 121-8.
• Yegutkin GG, Samburski SS, Jalkanen S. (2003) Soluble purine-converting enzymes circulate in human blood and regulate extracellular ATP level via counteracting pyrophosphatase and phosphotransfer reactions. FASEB J.; 17: 1328-30.
• Zimmermann H, Braun N. (1999) Ecto-nucleotidases: molecular structure, catalytic properties and functional roles in the nervous system. Prog Brain Res.; 120: 371-85.
• Zimmermann H. (2000) Extracellular metabolism of ATP and other nucleotides. Naunyn-Schmiedeberg's Arch Pharmacol.; 362: 299-309.