Characterization of Са2+,Мg2+-ATPase of blood lymphocytes in women with ovarian cancer

  • R. V. Barylyak Danylo Halytsky Lviv National Medical University
  • U. P. Iefremova Danylo Halytsky Lviv National Medical University
  • O. K. Onufrovych Danylo Halytsky Lviv National Medical University
  • O. V. Melnyk Danylo Halytsky Lviv National Medical University
  • D. Z. Vorobets Danylo Halytsky Lviv National Medical University
  • Z. D. Vorobets Danylo Halytsky Lviv National Medical University
Keywords: plasma membrane; Ca2 -ions; ATPase activity; ATP hydrolysis rate; affinity constant; Ca2 -pump; lymphocytes


Ionized Ca2+ is crucial for regulation of practically all intracellular processes, including tumor growth, cell proliferation, apoptosis, etc. The plasma membrane Ca2+,Mg2+-ATPase plays an important role in maintaining intracellular Ca2+ homeostasis. The function of this enzyme is to reduce the Ca2+ concentration in the cytosol, namely its transport against a concentration gradient in the extracellular medium. We have investigated the activity of plasma membrane Ca2+,Mg2+-ATPase of lymphocytes of practically (clinically) healthy women of different age groups and also patients with ovarian cancer stage III and IV. It was found that the enzyme activity in women of the older age group was not significantly reduced in relation to the activity of the younger age group. Thus, the value of the maximum rate of ATP hydrolysis by plasma membrane Ca2+,Mg2+-ATPase of blood lymphocytes in practically healthy women under the conditions of physiological norm was 1.1 times higher than under of pre-nosological state. In patients with ovarian cancer (stages III and IV), plasma membrane Ca2+,Mg2+-ATPase activity of blood lymphocytes significantly differed from the physiological norm and decreased by 1.6 and 1.8 times, compared with the physiological norm. The decrease of the plasma membrane Ca2+,Mg2+-ATPase activity of blood lymphocytes in patients with ovarian cancer indicates an increase of Ca2+ in the cytosol of lymphocytes. Determination of affinity constants showed that these values were in the submillimolar range of concentration, corresponding to the physiological concentration in the cell cytoplasm (0.5–5.0 mM). In healthy persons, under the condition of physiological norm, the affinity constant of plasma membrane Сa2+,Mg2+-ATPase to the ATP was 0.16 ± 0.02 mM and at pre-nosological state – 0.19 ± 0.02 mM. The affinity constant of plasma membrane Сa2+,Mg2+-ATPase of lymphocytes to ATP in patients with ovarian cancer (stage ІІІ) was 0.32 ± 0.03 mM and with ovarian cancer (stage IV) 0.35 ± 0.03 mM. That is, the affinity constant of plasma membrane Ca2+,Mg2+-ATPase of lymphocytes to ATP in patients with ovarian cancer was 2.0–2.1 times higher than this value for the blood lymphocytes in the control group (physiological norm). The kinetic analysis of Ca2+-activated, Mg2+-dependent hydrolysis of ATP in blood lymphocytes in women showed that the decrease in the activity of Ca2+,Mg2+-ATPase was due to a decrease in the affinity of the enzyme to the substrate (KATP increases 2-fold).


Agarwal, A., Mulgund, A., Hamada, A., & Chyatte, M. R. (2015). A unique view on male infertility around the globe. Reproductive Biology and Endocrinology, 13, 37.

Atig, F., Raffa, M., Habib, B. A., Kerkeni, A., Saad, A., & Ajina, M. (2012). Impact of seminal trace element and glutathione levels on semen quality of Tunisian infertile men. BioMed Central Urology, 12, 6.

Balercia, G., Moretti, S., Vignini, A., Magagnini, M., Mantero, F., Boscaro, M., Ricciardo-Lamonica, G., & Mazzanti, L. (2004). Role of nitric oxide concentrations on human sperm motility. Journal of Andrology, 25(2), 245–249.

Bonavida, B., Khineche, S., Huerta-Yepez, S., & Garban, H. (2006). Therapeutic potential of nitric oxide in cancer. Drug Resistance Updates, 9, 157–173.

Dillon, B. J., Holtsberg, F. W., Ensor, C. M., Bomalaski, J. S., & Clark, M. A. (2002). Biochemical characterization of the arginine degrading enzymes arginase and arginine deiminase and their effect on nitric oxide production. The Medical Science Monitor, 8(7), BR248–253.

Eskiocak, S., Gozen, A. S., Taskiran, A., Kilic, A. S., Eskiocak, M., Gulen, S. (2006). Effect of psychological stress on the L-arginine-nitric oxide pathway and semen quality. Brazilian Journal of Medical and Biological Research, 39(5), 581–588.

Fafula, R. V., Iefremova, U. P., Onufrovych, O. K., Maksymyuk, H. V., Besedina, A. S., Nakonechnyi, I. A., Vorobets, D. Z., & Vorobets, Z. D. (2018). Alterations in arginase-NO-synthase system of spermatozoa in human subjects with different fertility potential. Journal of Medical Biochemistry, 37, 1–7.

Fafula, R. V., Onufrovych, O. K., Iefremova, U. P., Vorobets, D. Z., & Vorobets, Z. D. (2016). Osoblyvosti arginaznogo shlyahu metabolismu L-argininu v spermatozoidach cholovikiv pry riznyh formach patospermiji [The peculiarrities of arginase pathway of L-arginine in spermatozoa in men with different forms of pathospermia]. Fiziologichnyj Zhurnal, 62(5), 83–90 (in Ukrainian).

Gür, S., & Kandemir, F. M. (2012). Relationships between seminal plasma arginase activity and spermatological parameters in rams. Andrologia, 44(2), 86–91.

Hadwan, M. H., Almashhedy, L. A., & Razzaq, A. S. (2014). Study of the effects of oral zinc supplementation on peroxynitrite levels, arginase activity and NO synthase activity in seminal plasma of Iraqi asthenospermic patients. Reproductive Biology and Endocrinology, 12, 1.

Jiang, J., & George, S. C. (2011). Modeling gas phase nitric oxide release in lung epithelial cells. Nitric Oxide, 25(3), 275–281.

Kosterin, S. A., & Burchinskaya, N. F. (1987). Metod opredileniya kineticheskih characteristic Ca2+-transportnyh sistem v subkletochnyh structurah gladkih myshc [Method for determination of kinetic parameters of Ca2+-transporting systems of subcellular structures of smooth muscle]. Ukrainian Biochemical Journal, 59(2), 66–69 (in Russian).

Kullisaar, T., Türk, S., Kilk, K., Ausmees, K., Punab, M., & Mandar, R. (2013). Increased levels of hydrogen peroxide and nitric oxide in male partners of infertile couples. Andrology, 1(6), 850–858.

Lowry, O. H., Rosebrough, N. J., Farr, A. L. (1951). Protein measurement with the Folin phenolreagent. The Journal of Biological Chemistry, 193, 265–275.

Luk, B. H., & Loke, A. Y. (2015). A review of supportive interventions targeting individuals or couples undergoing infertility treatment: Directions for the development of interventions. The Journal of Sex and Marital Therapy, 10, 1–19.

Mori, M. (2007). Regulation of nitric oxide synthesis and apoptosis by arginase and arginine recycling. The Journal of Nutrition, 137, 1616–1620.

Porro, B., Eligini, S., Squellerio, I., Tremoli, E., & Cavalca, V. (2014). The red blood cell: A new key player in cardiovascular homoeostasis? Focus on the nitric oxide pathway. Biochemical Society Transactions, 42(4), 996–1000.

Racke, K., & Warnken, M. (2010). L-arginine metabolic pathways. The Open Nitric Oxide Journal, 2, 9–19.

Shen, L. J., Beloussow, K., & Shen, W. C. (2005). Accessibility of endothelial and inducible nitric oxide synthase to the intracellular citrulline–arginine regeneration pathway. Biochemical Pharmacology, 69, 97–104.

Stephen, D. C., Hong, Y., Wayne, W. G., Rita, M. K., Paul, Y., & Ramaswamy, K. I. (2004). Arginases I and II: Do their functions overlap? Molecular Genetics and Metabolism, 81, 38–44.

Topal, G., Brunet, A., Walch, L., Boucher, J. L., & David-Dufilho, M. (2006) Mitochondrial arginase II modulates nitric-oxide synthesis through nonfreely exchangeable L-arginine pools in human endothelial cells. The Journal of Pharmacology and Experimental Therapeutics, 318, 1368–1374.

Türk, G., Güra, S., Kandemirb, F. M., & Sünmez, M. (2011). Relationship between seminal plasma arginase activity and semen quality in Saanen bucks. Small Ruminant Research, 97(1–3), 83–87.

Vanhoutte, P. M. (2008). Arginine and arginase: Endothelial NO synthase double crossed? Circulation Research, 102, 866–868.

Venkatakrishnan, P., Nakayasu, E. S., Almeida, I. C., & Miller, R. (2010). Arginase activity in mitochondria – An interfering factor in nitric oxide synthase activity assays. Biochemical and Biophysical Research Communications, 394(3), 448–452.

Winters, B. R., & Walsh, T. J. (2014). The epidemiology of male infertility. Urologic Clinics of North Americ, 41(1), 195–204.


How to Cite
Barylyak, R. V., Iefremova, U. P., Onufrovych, O. K., Melnyk, O. V., Vorobets, D. Z., & Vorobets, Z. D. (2018). Characterization of Са2+,Мg2+-ATPase of blood lymphocytes in women with ovarian cancer. Regulatory Mechanisms in Biosystems, 9(1), 85-89.

Most read articles by the same author(s)