ATP and ADP hydrolysis in cell membranes from rat myometrium
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Milošević, MajaPetrovic, Snjezana
Velickovic, Natasa
Grković, Ivana
Ignjatović, Marija
Horvat, Anica
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Extracellular nucleotides affect female reproductive functions, fertilization, and pregnancy. The aim of this study was to investigate biochemical characteristics of ATP and ADP hydrolysis and identify E-NTPDases in myometrial cell membranes from Wistar albino rats. The apparent K (m) values were 506.4 +/- A 62.1 and 638.8 +/- A 31.3 mu M, with a calculated V (max) (app) of 3,973.0 +/- A 279.5 and 2,853.9 +/- A 79.8 nmol/min/mg for ATP and ADP, respectively. The enzyme activity described here has common properties characteristic for NTPDases: divalent cation dependence; alkaline pH optimum for both substrates, insensitivity to some of classical ATPase inhibitors (ouabain, oligomycine, theophylline, levamisole) and significant inhibition by suramine and high concentration of sodium azides (5 mM). According to similar apparent K-m values for both substrates, the ATP/ADP hydrolysis ratio, and Chevillard competition plot, NTPDase1 is dominant ATP/ADP hydrolyzing enzyme in myometrial cell m...embranes. RT-PCR analysis revealed expression of three members of ectonucleoside triphosphate diphosphohydrolase family (NTPDase 1, 2, and 8) in rat uterus. These findings may further elucidate the role of NTPDases and ATP in reproductive physiology.
Keywords:
ATP/ADP hydrolysis / NTPDase / Purinergic signaling / Uterus / RatSource:
Molecular and Cellular Biochemistry, 2012, 371, 1-2, 199-208Funding / projects:
- Molecular mechanisms of cellular responses on pathological changes in central neuronal system and peripheral organs of mammals (RS-MESTD-Basic Research (BR or ON)-173044)
- Cellular and molecular basis of neuroinflamation: potential targets for translational medicine and therapy (RS-MESTD-Integrated and Interdisciplinary Research (IIR or III)-41014)
DOI: 10.1007/s11010-012-1436-2
ISSN: 0300-8177; 1573-4919
PubMed: 22956447
WoS: 000310425100020
Scopus: 2-s2.0-84868576006
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VinčaTY - JOUR AU - Milošević, Maja AU - Petrovic, Snjezana AU - Velickovic, Natasa AU - Grković, Ivana AU - Ignjatović, Marija AU - Horvat, Anica PY - 2012 UR - https://vinar.vin.bg.ac.rs/handle/123456789/5131 AB - Extracellular nucleotides affect female reproductive functions, fertilization, and pregnancy. The aim of this study was to investigate biochemical characteristics of ATP and ADP hydrolysis and identify E-NTPDases in myometrial cell membranes from Wistar albino rats. The apparent K (m) values were 506.4 +/- A 62.1 and 638.8 +/- A 31.3 mu M, with a calculated V (max) (app) of 3,973.0 +/- A 279.5 and 2,853.9 +/- A 79.8 nmol/min/mg for ATP and ADP, respectively. The enzyme activity described here has common properties characteristic for NTPDases: divalent cation dependence; alkaline pH optimum for both substrates, insensitivity to some of classical ATPase inhibitors (ouabain, oligomycine, theophylline, levamisole) and significant inhibition by suramine and high concentration of sodium azides (5 mM). According to similar apparent K-m values for both substrates, the ATP/ADP hydrolysis ratio, and Chevillard competition plot, NTPDase1 is dominant ATP/ADP hydrolyzing enzyme in myometrial cell membranes. RT-PCR analysis revealed expression of three members of ectonucleoside triphosphate diphosphohydrolase family (NTPDase 1, 2, and 8) in rat uterus. These findings may further elucidate the role of NTPDases and ATP in reproductive physiology. T2 - Molecular and Cellular Biochemistry T1 - ATP and ADP hydrolysis in cell membranes from rat myometrium VL - 371 IS - 1-2 SP - 199 EP - 208 DO - 10.1007/s11010-012-1436-2 ER -
@article{ author = "Milošević, Maja and Petrovic, Snjezana and Velickovic, Natasa and Grković, Ivana and Ignjatović, Marija and Horvat, Anica", year = "2012", abstract = "Extracellular nucleotides affect female reproductive functions, fertilization, and pregnancy. The aim of this study was to investigate biochemical characteristics of ATP and ADP hydrolysis and identify E-NTPDases in myometrial cell membranes from Wistar albino rats. The apparent K (m) values were 506.4 +/- A 62.1 and 638.8 +/- A 31.3 mu M, with a calculated V (max) (app) of 3,973.0 +/- A 279.5 and 2,853.9 +/- A 79.8 nmol/min/mg for ATP and ADP, respectively. The enzyme activity described here has common properties characteristic for NTPDases: divalent cation dependence; alkaline pH optimum for both substrates, insensitivity to some of classical ATPase inhibitors (ouabain, oligomycine, theophylline, levamisole) and significant inhibition by suramine and high concentration of sodium azides (5 mM). According to similar apparent K-m values for both substrates, the ATP/ADP hydrolysis ratio, and Chevillard competition plot, NTPDase1 is dominant ATP/ADP hydrolyzing enzyme in myometrial cell membranes. RT-PCR analysis revealed expression of three members of ectonucleoside triphosphate diphosphohydrolase family (NTPDase 1, 2, and 8) in rat uterus. These findings may further elucidate the role of NTPDases and ATP in reproductive physiology.", journal = "Molecular and Cellular Biochemistry", title = "ATP and ADP hydrolysis in cell membranes from rat myometrium", volume = "371", number = "1-2", pages = "199-208", doi = "10.1007/s11010-012-1436-2" }
Milošević, M., Petrovic, S., Velickovic, N., Grković, I., Ignjatović, M.,& Horvat, A.. (2012). ATP and ADP hydrolysis in cell membranes from rat myometrium. in Molecular and Cellular Biochemistry, 371(1-2), 199-208. https://doi.org/10.1007/s11010-012-1436-2
Milošević M, Petrovic S, Velickovic N, Grković I, Ignjatović M, Horvat A. ATP and ADP hydrolysis in cell membranes from rat myometrium. in Molecular and Cellular Biochemistry. 2012;371(1-2):199-208. doi:10.1007/s11010-012-1436-2 .
Milošević, Maja, Petrovic, Snjezana, Velickovic, Natasa, Grković, Ivana, Ignjatović, Marija, Horvat, Anica, "ATP and ADP hydrolysis in cell membranes from rat myometrium" in Molecular and Cellular Biochemistry, 371, no. 1-2 (2012):199-208, https://doi.org/10.1007/s11010-012-1436-2 . .