Characterization of ectonucleoside triphosphate diphosphohydrolase (E-NTPDase; EC 3.6.1.5) activity in mouse peritoneal cavity cells
| Autor: | Daniela B.R. Leal, Dhébora Albuquerque Dias, Lucas Derbocio dos Santos, Pedro Mendes dos Santos, Maria Rosa Chitolina Schetinger, Jeandre Augusto dos Santos Jaques, Bruna de Barros Penteado, Carla Cardozo Pinto de Arruda |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Cell Survival Neutrophils ATPase Clinical Biochemistry Biochemistry Substrate Specificity Mice 03 medical and health sciences chemistry.chemical_compound Peritoneal cavity 0302 clinical medicine Adenine nucleotide Cations Sodium fluoride medicine Animals Magnesium Nucleotide Lymphocytes Peritoneal Cavity Cells Cultured Adenosine Triphosphatases chemistry.chemical_classification Mice Inbred BALB C biology Chemistry Apyrase Macrophages Temperature Cell Biology General Medicine Hydrogen-Ion Concentration Molecular biology Kinetics Adenosine diphosphate 030104 developmental biology medicine.anatomical_structure 030220 oncology & carcinogenesis biology.protein Calcium Female Adenosine triphosphate |
| Zdroj: | Cell Biochemistry and Function. 35:358-363 |
| ISSN: | 0263-6484 |
| DOI: | 10.1002/cbf.3281 |
| Popis: | This study aimed to characterize the activity of ectonucleoside triphosphate diphosphohydrolase (E-NTPDase; EC 3.6.1.5) in peritoneal cavity cells from BALB/c mice. E-NTPDase was activated in the presence of both calcium (1.5mM) and magnesium (1.5mM) ions. However, the activity was higher in the presence of Ca2+ . A pH of 8.5 and temperature of 37°C were the optimum conditions for catalysis. The apparent Km values were 0.51mM and 0.66mM for the hydrolysis of adenosine triphosphate (ATP) and adenosine diphosphate (ADP), respectively. The Vmax values were 136.4 and 120.8 nmol Pi/min/mg of protein for ATPase and ADPase activity, respectively. Nucleotide hydrolysis was inhibited in the presence of sodium azide (20mM, ATP: P < .05; ADP: P < .001), sodium fluoride (20mM; ATP and ADP: P < .001), and suramin (0.3mM; ATP: P < .01; ADP: P < .05), which is a known profile for NTPDase inhibition. Although all of the diphosphate and triphosphate nucleotides that were tested were hydrolyzed, enzyme activity was increased when adenine nucleotides were used as substrates. Finally, we stress that knowledge of the E-NTPDase catalytic biochemical properties in mouse peritoneal cavity cells is indispensable for properly determining its activity, as well as to fully understand the immune response profile in both healthy and sick cells. |
| Databáze: | OpenAIRE |
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