Summary
We examined the interactions of cAMP-dependent protein kinase and varying aqueous Cl− concentrations in modulating the activity of Cl− channels obtained by fusing basolaterally enriched renal outer medullary vesicles into planar lipid bilayers. Under the present experimental conditions, thecis andtrans solutions face the extracellular and intracellular aspects of these Cl− channels, respectively. Raising thetrans Cl− concentration from 2 to 50mm increased the channel open-time probability, raised the unit channel conductance, and affected the voltage-independent determinant (ΔG) of channel activity but not the gating charge (Winters, C.J., Reeves, W.B., Andreoli, T.E. 1990.J. Membrane Biol. 118:269–278). With 2mm trans KCl,trans addition of the catalytic subunit of PKA (C-PKA) plus ATP increased channel open-time probability and altered the voltage-independent determinant of channel activity without affecting either unit channel conductance or gating charge. The effect was ATP specific, did not occur with (C-PKA plus ATP) addition tocis solutions, and was abolished by denaturing C-PKA. Finally, (C-PKA plus ATP) activation of channel activity was not detected with relatively high (50mm)trans Cl− concentrations. These data indicate that (C-PKA plus ATP) might modulate Cl− channel activity by phosphorylation at or near the Cl−-sensitive site on the intracellular face of these channels.
Similar content being viewed by others
References
Bayliss, J.M., Reeves, W.B., Andreoli, T.E. 1990. Cl− transport in basolateral renal medullary vesicles: I. Cl− transport in intact vesicles.J. Membrane Biol. 113:49–56
Burnham, C., Karlish, S.J.D., Jorgensen, P.L. 1985. Identification and reconstitution of a Na+/K+/Cl− cotransporter and K+ channel from luninal membranes of renal red outer medulla.Biochim. Biophys. Acta 821:461–469
Geck, P., Pfeiffer, B. 1985. Na+, K+, 2Cl cotransport in animal cells—its role in volume regulation.Ann. NY Acad. Sci. 456:166–182
Greger, R., Oberleithner, H., Schlatter, E., Cassola, A.C., Weidtke, C. 1983. Chloride activity in cells of isolated perfused cortical thick ascending limbs of rabbit kidney.Pfluegers Arch. 399:29–34
Halm, D.R., Rechkemmer, G.R., Schoumacher, R.A., Frizzell, R.A. 1988. Apical membrane chloride channels in a colonic cell line activated by secretory agonists.Am. J. Physiol. 254:C505-C511
Hebert, S.C., Andreoli, T.E. 1984. Effects of antidiuretic hormone on cellular conductive pathways in mouse medullary thick ascending limbs of Henle: II. Determinants of the ADH-mediated increases in transepithelial voltage and in net Cl− absorption.J. Membrane Biol. 80:221–233
Hebert, S.C., Culpepper, R.M., Andreoli, T.E. 1981. NaCl transport in mouse medullary thick ascending limbs. I. Functional nephron heterogeneity and ADH-stimulated NaCl cotransport.Am. J. Physiol. 241:F412-F431
Hebert, S.C., Culpepper, R.M., Andreoli, T.E. 1981. NaCl transport in mouse medullary thick ascending limbs. II. ADH enhancement of transcellular NaCl co-transport: origin of transepithelial voltage.Am. J. Physiol. 241:F432-F442
Hebert, S.C., Friedman, P.A., Andreoli, T.E. 1984. Effects of antidiuretic hormone on cellular conductive pathways in mouse medullary thick ascending limbs of Henle: I. ADH increases transcellular conductance pathways.J. Membrane Biol. 80:201–219
Kinne, R., Kinne-Saffran, E., Scholermann, B., Schultz, H. 1986. The anion specificity of the sodium-potassium-chloride cotransporter in rabbit kidney outer medulla: Studies on medullary plasma membranes.Pfluegers Arch. 407:S168-S173
Li, M., McCann, J.D., Liedtke, C.M., Nairn, A.C., Greengard, P., Welsh, M.J. 1988. Cyclic AMP-dependent protein kinase opens chloride channels in normal but not cystic fibrosis airway epithelium.Nature 331:358–360
Mandel, K.G., Dharmsathaphorn, K., McRoberts, J.A. 1986. Characterization of a cyclic-AMP-activated potassium transport in a human colonic epithelial cell line.J. Biol. Chem. 261:704–712
Molony, D.A., Reeves, W.B., Hebert, S.C., Andreoli, T.E.. 1987. ADH increases apical Na+,K+,2Cl− entry in mouse medullary thick ascending limbs of Henle.Am. J. Physiol. 252:F177-F187.
Paulais, M., Teulon, J. 1990. cAMP-activated chloride channel in the basolateral membrane of the thick ascending limb of the mouse kidney.J. Membrane Biol. 113:253–260
Reeves, W.B., Andreoli, T.E. 1990. Cl− transport in basolateral renal medullary vesicles: II. Cl− channels in planar lipid bilayers.J. Membrane Biol. 113:57–65
Reeves, W.B., McDonald, G.A., Mehta, P., Andreoli, T.E. 1989. Activation of K+ channels in renal medullary vesicles by cAMP-dependent protein kinase.J. Membrane Biol. 109:65–72
Reimann, E.M., Beham, R.A. 1983. Catalytic subunit of cAMP-dependent protein kinase.Methods Enzymol. 99:51–55
Schlatter, E., Greger, R. 1985. cAMP increases the basolateral Cl− conductance in the isolated perfused medullary thick ascending limb of Henle's loop of the mouse.Pfluegers Arch. 405:367–376
Schlender, K.K., Reiman, E.R. 1977. Glycogen synthase kinases.J. Biol. Chem. 252:2384–2389
Schoumacher, R.A., Shoemaker, R.L., Halm, D.R., Tallant, E.A., Wallace, R.W., Frizzell, R.A. 1987. Phosphorylation fails to activate chloride channels from cystic fibrosis airway cells.Nature 330:752–754
Valdivia, H.H., Dubinsky, W.P., Coronado, R. 1988. Reconstitution and phosphorylation of chloride channels from airway epithelium membranes.Science 242:1441–1444
Winters, C.J., Reeves, W.B., Andreoli, T.E. 1990. cAMP-dependent protein kinase (PKA) and Cl− independently activate intracellular faces of Cl− channels in basolateral medullary vesicles.J. Am. Soc. Nephrol. 1:695
Winters, C.J., Reeves, W.B., Andreoli, T.E. 1990. Cl− channels in basolateral renal medullary membranes. III. Determinants of single-channel activity.J. Membrane Biol. 118:269–278
Wittner, M., deStefano, A., Wangemann, P., Nitschke, R., Greger, R., Bailly, C., Amiel, C., Roinel, N., deRouffignac, C. 1988. Differential effects of ADH on sodium, chloride, potassium, calcium and magnesium transport in cortical and medullary thick ascending limbs of mouse nephron.Pfluegers Arch. 412:516–523
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Winters, C.J., Reeves, W.B. & Andreoli, T.E. Cl− channels in basolateral renal medullary membrane vesicles: IV. Analogous channel activation by Cl− or cAMP-dependent protein kinase. J. Membrain Biol. 122, 89–95 (1991). https://doi.org/10.1007/BF01872742
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01872742