Abstract
The possible correlation between Pglycoprotein (PGP) and volume-sensitive Cl− channel was examined in a pair of cell lines: a subline of the human epidermoid KB cell (KB-3-1) and the corresponding MDR1-transfected cell line (KB-G2). Western blot analysis and indirect immunofluorescence studies indicated that KB-G2, but not KB-3-1, exhibits the PGP expression. Patch-clamp whole-cell recordings showed that osmotic swelling activates Cl− currents not only in PGP-expressing but also in PGP-lacking cells. The amplitude of the maximal current was indistinguishable between both cells. Activation of protein kinase C (PKC) or loading with a PKC inhibitor failed to affect the swelling-induced activation of the Cl− currents in both cells. The relation between whole-cell Cl− currents and cell size measured simultaneously showed that volume sensitivity of the Cl− channel was augmented by the PGP expression irrespective of the activity of PKC on the plasma membrane. A similar increase in volume sensitivity of the Cl− channel was also induced by the expression of the ATP hydrolysis-deficient PGP mutant, K433M. We conclude that P-glycoprotein does not represent the volume-sensitive Cl− channel but that its expression modulates volume sensitivity of the Cl− channel in a manner independent of its ATPase activity or of the protein kinase C activity.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Akiyama, S.-I., Fojo, A., Hanover, J.A., Pastan, I., Gottesman, M.M. 1985. Isolation and genetic characterization of human KB cell lines resistant to multiple drugs. Somat. Cell. Mol. Gen. 11:117–126
Altenberg, G.A., Vanoye, C.G., Han, E.S., Deitmer, J.W., Reuss, L. 1994. Relationships between rhodamine 123 transport, cell volume, and ion-channel function of P-glycoprotein. J. Biol. Chem. 269:7145–7149
Borst, P., Schinkel, A.H., Smit, J.J.M., Wagenaar, E., Van Deemter, L., Smith, A.J., Eijdems, E.W.H.M., Baas, F., Zaman, G.J.R. 1994. Classical and novel forms of multidrug resistance and the physiological functions of P-glycoproteins in mammals. Phamac. Ther. 60:289–299
Chambers, T.C., McAvoy, E.M., Jacobs, J.W., Eilon, G. 1990. Protein kinase C phosphorylates P-glycoprotein in multidrug resistant human KB carcinoma cells. J. Biol. Chem. 265:7679–7686
Chan, H.C., Fu, W.O., Chung, Y.W., Huang, S.J., Chan, P.S.F., Wong, P.Y.D. 1994. Swelling-induced anion and cation conductances in human epididymal cells. J. Physiol. 478:449–460
Coca-Prados, M., Sánchez-Torres, J., Peterson-Yantorno, K., Civan, M.M. 1996. Association of C1C-3 channel with Cl− transport by human nonpigmented ciliary epithelial cells. J. Membrane Biol. 150:197–208
De Greef, C., van der Heyden, S., Viana, F., Eggermont, J., De Bruijn, E.A., Raeymaekers, L., Droogmans, G., Nilius, B. 1995. Volume-activated chloride currents are not correlated with P-glycoprotein expression. Pfluegers Arch. 430:296–298
Dong, Y., Chen, G., Durán, G.E., Kouyama, K., Chao, A.C., Sikic, B.I., Gollapudi, S.V., Gupta, S., Gardner, P. 1994. Volume-activated chloride current is not related to P-glycoprotein overexpression. Cancer Res. 54:5029–5032
Ehring, G.R., Osipchuk, Y.V., Cahalan, M.D. 1994. Swelling-activated chloride channels in multidrug-sensitive and -resistant cells. J. Gen. Physiol. 104:1129–1161
Gill, D.R., Hyde, S.C., Higgins, C.F., Valverde, M.A., Mintenig, G.M., Sepúlveda, F.V. 1992. Separation of drug transport and chloride channel functions of the human multidrug resistance Pglycoprotein. Cell 71:23–32
Gschwentner, M., Nagel, U.O., Wöll, E., Schmarda, A., Ritter, M., Paulmichl, M. 1995. Antisense oligonucleotides suppress cell-volume-induced activation of chloride channels. Pfluegers Arch. 430:464–470
Hardy, S.P., Goodfellow, H.R., Valverde, M.A., Gill, D.R., Sepúlveda, F., Higgins, CF. 1995. Protein kinase C-mediated phosphorylation of the human multidrug resistance P-glycoprotein regulates cell volume-activated chloride channels. EMBO J. 14:68–75
Higgins, C.F. 1992. ABC transporters: From microorganisms to man. Ann. Rev. Cell. Biol. 8:67–113
Kikkawa, U., Minakuchi, R., Takai, Y., Nishizuka, Y. 1983. Calcium-activated, phospholipid-dependent protein kinase (protein kinase C) from rat brain. Methods Enzymol. 99:288–298
Kubo, M., Okada, Y. 1992. Volume-regulatory Cl− channel currents in cultured human epithelial cells. J. Physiol. 456:351–371
Loo, T.W., Clarke, D.M. 1995. Rapid purification of human Pglycoprotein mutants expressed transiently in HEK 293 cells by nickel-chelate chromatography and characterization of their drugstimulated ATPase activities. J. Biol. Chem. 270:21449–21452
Luckie, D.B., Krouse, M.E., Harper, K.L., Law, T.C., Wine, J.J. 1994. Selection for MDR1/P-glycoprotein enhances swelling-activated K+ and Cl− currents in NIH/3T3 cells. Am. J. Physiol. 267:C650-C658
Ma, L., Marquardt, D., Takemoto, L., Center, M.S. 1991. Analysis of P-glycoprotein phosphorylation in HL60 cells isolated for resistance to vincristine. J. Biol. Chem. 266:5593–5599
Nilius, B., Sehrer, J., Viana, F., De Greef, C., Raeymaekers, L., Eggermont, J., Droogmans, G. 1994. Volume-activated Cl− currents in different mammalian nonexcitable cell types. Pfluegers Arch. 428:364–371
Nishizuka, Y. 1984. The role of protein kinase C in cell surface transduction and tumour promotion. Nature 308:693–698
Nishizuka, Y. 1986. Studies and perspectives of protein kinase C. Science 233:305–312
Posada, J., Vichi, P., Tritton, T.R. 1989. Protein kinase C in adriamycin action and resistance in mouse sarcoma 180 cells. Cancer Res. 49:6634–6639
Rasola, A., Galietta, L.J., Gruenert, D.C., Romeo, G. 1994. Volume-sensitive chloride currents in four epithelial cell lines are not directly correlated to the expression of the MDR-1 gene. J. Biol. Chem. 269:1432–1436
Strange, K., Emma, F., Jackson, P.S. 1996. Cellular and molecular physiology of volume-sensitive anion channels. Am. J. Physiol. 270:C711-C730
Tominaga, M., Tominaga, T., Miwa, A., Okada, Y. 1995. Volume-sensitive chloride channel activity does not depend on endogenous P-glycoprotein. J. Biol. Chem. 270:27887–27893
Ueda, K., Okamura, N., Hirai, M., Tanigawara, Y., Saeki, T., Kioka, N., Komano, T., Hori, R. 1992. Human P-glycoprotein transports cortisol, aldosterone, and dexamethasone, but not progesterone. J. Biol. Chem. 267:24248–24252
Ueda, K., Shimabuku, A.M., Konishi, H., Fujii, Y., Takabe, S., Nishi, K., Yoshida, M., Beppu, T., Komano, T. 1993. Functional expression of human P-glycoprotein in Schizosaccharomyces pombe. FEBS Lett. 330:279–282
Valverde, M.A., Bond, T.D., Hardy, S.P., Taylor, J.C., Higgins, C.F., Altamirano, J., Alvarez-Leefmans, F.J. 1996. The multidrug resistance P-glycoprotein modulates cell regulatory volume decrease. EMBO J. 15:4460–4468
Valverde, M.A., Díaz, M., Sepúlveda, F.V., Gill, D.R., Hyde, S.C., Higgins, C.F. 1992. Volume-regulated chloride channels associated with the human multidrug-resistance P-glycoprotein. Nature 355:830–833
Viana, F., Van Acker, K., De Greef, C., Eggermont, J., Raeymaekers, L., Droogmans, G., Nilius, B. 1995. Drug-transport and volume-activated chloride channel functions in human erythroleukemia cells: Relation to expression level of P-glycoprotein. J. Membrane Biol. 145:87–98
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Miwa, A., Ueda, K. & Okada, Y. Protein kinase C-independent correlation between P-glycoprotein expression and volume sensitivity of Cl− channel. J. Membrane Biol. 157, 63–69 (1997). https://doi.org/10.1007/s002329900216
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/s002329900216