Abstract
The present study aimed to find out a link between ageing of rat and lamprey erythrocytes and activity of two isoforms of protein kinase C (PKC), РKСα and РKСζ. The whole cell population was separated into fractions of different ages in Percoll density gradient. The validity of separation was confirmed by the number of immature erythrocytes, reticulocytes. PKC activity was analyzed in cytosolic and membrane cell fractions. Rat erythrocytes express both PKC isoforms, РKСα and РKСζ, whereas lamprey erythrocytes express only РKСζ. РKСα is identified as a major band at ~ 80 kDa and minor bands at ~ 55–65 kDa; РKСζ is represented by a single band at ~ 80 kDa. In young rat erythrocytes, РKСα is detected mainly in cytosolic fractions, while in membrane fractions its level is by far lower. As cells age, PKCα is translocated from the cytosol to membranes and undergoes proteolytic degradation due to repeated cycles of activation. As a result, in aged erythrocytes relative total PKCα expression (as a sum of expressions in the cytosol and membranes per total protein level) diminishes, indicating a depletion of the PKCα pool and a decline in its functional activity. In both animal species, a highest PKCζ level is observed in the cytosol of young erythrocytes. Erythrocyte ageing is accompanied by a gradual decrease in expression of free cytosolic PKCζ and concurrent increase in the level of its membrane-bound forms. However, in contrast to PKCα, PKCζ is not proteolyzed; its total level in cells and perhaps functional activity do not change throughout the erythrocyte lifespan.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Gusev, G.P., Govekar, R., Gadewal, N., and Agalakova, N.I., Understanding quasi-apoptosis of the most numerous enucleated components of blood needs detailed molecular autopsy, Ageing Res. Rev., 2017, vol. 35, pp. 46–62.
Li, H., Feng, L., Jiang, W., Liu, Y., Jiang, J., Zhang, Y., Wu, P., and Zhou, X., Ca2+ and caspases are involved in hydroxyl radical-induced apoptosis in erythrocytes of Jian carp (Cyprinus caprio var. Jian), Fish Physiol. Biochem., 2015, vol. 41, pp. 1305–1319.
Bratosin, D., Estaquier, J., Slomianny, C., Tissier, J.P., Quatannens, B., Bulai, T., Mitrofan, L., Marinescu, A., Trandaburu, I., Ameisen, J.C., and Montreuil, J., On the evolution of erythrocyte programmed cell death: apoptosis of Rana esculenta nucleated red blood cells involves cysteine proteinase activation and mitochondrion permeabilization, Biochimie, 2004, vol. 86, pp. 183–192.
Weil, M., Jacobson, M.D., and Raff, M.C., Are caspases involved in the death of cells with a transcriptionally inactive nucleus? Sperm and chicken erythrocytes, J. Cell Sci., 1998, vol. 111, pp. 2707–2715.
Newton, A.C., Protein kinase C: poised to signal, Am. J. Physiol. Endocrinol. Metab., 2010, vol. 298, pp. E395–E402.
Reyland, M.E. and Bradford, A.P., PKC and the control of apoptosis, Protein Kinase C in Cancer Signaling and Therapy, Kazanietz, M.G., Ed., Springer, 2010, pp. 189–222.
Linch, M., Riou, P., Claus, J., Cameron, A.J., de Naurois, J., Larijani, B., Ng, T., McDonald, N.Q., and Parker, P.J., Functional implications of assigned, assumed and assembled PKC structures, Biochem. Soc. Trans., 2014, vol. 42, pp. 35–41.
Govekar, R.B. and Zingde, S.M., Protein kinase C isoforms in human erythrocytes, Ann. Hematol., 2001, vol. 80, pp. 531–534.
Agalakova, N.I. and Gusev, G.P., Transient activation of protein kinase C contributes to fluorideinduced apoptosis of rat erythrocytes, Toxicol. in Vitro, 2013, vol. 27, pp. 2335–2341.
Rennie, C.M., Thompson, S., Parker, A.C., and Maddy, A., Human erythrocyte fraction in “Percoll” density gradients, Clin. Chim. Acta, 1979, vol. 98, pp. 119–125.
Agalakova, N.I., Ivanova, T.I., Gusev, G.P., Nazarenkova, A.V., and Sufiyeva, D.A., Apoptotic death in erythrocytes of lamprey Lampetra fluviatilis induced by ionomycin and tert-butyl hydroperoxide, Comp. Biochem. Physiol. C, 2017, vol. 194, pp. 48–60.
Fathallah, H., Sauvage, M., Romero, J.R., Canessa, M., and Giraud, F., Effects of PKC alpha activation on Ca2+ pump and K(Ca) channel in deoxygenated sickle cells, Am. J. Physiol., 1997, vol. 273, pp. C1206–1214.
Ai, Z. and Cohen, C.M., Phorbol 12-myristate 13-acetate-stimulated phosphorylation of erythrocyte membrane skeletal proteins is blocked by calpain inhibitors: possible role of protein kinase M, Biochem. J., 1993, vol. 296, pp. 675–683.
Jindal, H.K., Ai, Z., Gascard, P., Horton, C., and Cohen, C.M., Specific loss of protein kinase activities in senescent erythrocytes, Blood, 1996, vol. 88, pp. 1479–1487.
Postnov, Y.V., Kravtsov, G.M., Orlov, S.N., Pokudin, N.I., Postnov, I.Y., and Kotelevtsev, Y.V., Effect of protein kinase C activation on cytoskeleton and cation transport in human erythrocytes. Reproduction of some membrane abnormalities revealed in essential hypertension, Hypertension, 1988, vol. 12, pp. 267–273.
Andrews, D. A., Yang, L., and Low, P.S., Phorbol ester stimulates a protein kinase C-mediated agatoxin-TK-sensitive calcium permeability pathways in human red blood cells, Blood, 2002, vol. 100, pp. 3392–3399.
Kowluru, R., Bitensky, M.W., Kowluru, A., Dembo, M., Keaton, P.A., and Buican, T., Reversible sodium pump defect and swelling in the diabetic rat erythrocyte: effects on filterability and implications, Proc. Natl. Acad. Sci. USA, 1989, vol. 86, pp. 3327–3331.
Belloni-Olivi, L., Annadata, M., Goldstein, G.W., and Bressler, J., Phosphorylation of membrane proteins in erythrocytes treated with lead, Biochem. J., 1996, vol. 315, pp. 401–406.
Govekar, R.B., Kawle, P.D., Advani, S.H., and Zingde, S.M., Reduced PKCa activity induces senescent phenotype in erythrocytes, Anemia, 2012, ID 168050.
Sauvage, M., Maziere, P., Fathallah, H., and Giraud, F., Insulin stimulates NHE1 activity by sequential activation of phosphatidylinositol 3-kinase and protein kinase Cζ in human erythrocytes, Eur. J. Biochem., 2000, vol. 267, pp. 955–962.
Manno, S., Takakuwa, Y., and Mohandas, N., Modulation of erythrocyte membrane mechanical function by protein 4.1 phosphorylation, J. Biol. Chem., 2005, vol. 280, pp. 7581–7587.
Author information
Authors and Affiliations
Corresponding author
Additional information
Original Russian Text © N.I. Agalakova, I.A. Khvorova, T.I. Ivanova, 2018, published in Zhurnal Evolyutsionnoi Biokhimii i Fiziologii, 2018, Vol. 54, No. 3, pp. 154–162.
Rights and permissions
About this article
Cite this article
Agalakova, N.I., Khvorova, I.A. & Ivanova, T.I. Comparative Analysis of PKСα and PKCζ Activities in Rat and Lamprey Erythrocytes of Different Ages. J Evol Biochem Phys 54, 175–184 (2018). https://doi.org/10.1134/S002209301803002X
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S002209301803002X