Skip to main content
Log in

Patch-clamp studies on the anomalous mole fraction effect of the K+ channel in cytoplasmic droplets ofNitella: An attempt to distinguish between a multi-ion single-file pore and an enzyme kinetic model with lazy state

  • Articles
  • Published:
The Journal of Membrane Biology Aims and scope Submit manuscript

Summary

Patch-clamp studies have been employed in order to check whether the assumption of a multi-ion single-file pore is necessary for the explanation of the anomalous mole fraction effect or whether this effect can also be explained by a single-barrier enzyme kinetic model. Experiments in the cell-attached configuration were done on the tonoplast membrane of cytoplasmic droplets ofNitella in solutions containing 150 mol m−3 of K+ plus Tl+ with seven different K+/Tl+ ratios. At first sight, the results seem to support the multi-ion single-file pore, because apparent open channel conductivity displays the anomalous mole fraction effect, whereas open-probability has not been found to be dependent on the K+/Tl+ ratio. Changes in open probability would be expected for a single-barrier enzyme kinetic model with a lazy state. On the other hand, the lazy-state model is more successful in explaining the measuredI-V curves. The entire slope of the apparent open channel current-voltage curves rotates with changing K+/Tl+ ratios in the whole voltage range between −100 and + 80 mV. Numerical calculations on the basis of multi-ion single-file pores could create the anomalous mole fraction effect only in a limited voltage range with intersectingI-V curves. The apparent absence of an effect on open probability which is postulated by the lazy-state model can be explained if switching into and out of the lazy state is faster than can be resolved by the temporal resolution of 1 msec.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bertl, A. 1989. Current-voltage relationships of a sodium-sensitive potassium channel in the tonoplast ofChara corallina.J. Membrane Biol. 109:9–19

    Article  Google Scholar 

  • Bertl, A., Gradmann, D. 1987. Current-voltage relationships of potassium channels in the plasmalemma ofAcetabularia.J. Membrane Biol. 99:41–49

    Article  Google Scholar 

  • Bertl, A., Klieber, H.G., Gradmann, D. 1988. Slow kinetics of a potassium channel inAcetabularia.J. Membrane Biol. 102:141–152

    Article  Google Scholar 

  • Bezanilla, F. 1985. A high capacity data recording device based on a digital audio processor and a video cassette recorder.Biophys. J. 47:437–441

    PubMed  Google Scholar 

  • Blatt, M.R. 1988. Potassium-dependent, bipolar gating of K+ channels in guard cells.J. Membrane Biol. 102, 235–246

    Article  Google Scholar 

  • Fisahn, J., Hansen, U.-P., Gradmann, D. 1986. Determination of charge, stoichiometry and reaction constants fromI-V curve studies on a K+ transporter inNitella.J. Membrane Biol. 94:245–252

    Article  Google Scholar 

  • Goodwin, G.C., Sin, K.S. 1984. Adaptive Filtering Prediction and Control. pp. 505–507, Prentice-Hall, Englewood Cliffs

    Google Scholar 

  • Gradmann, D., Klieber, H.G., Hansen, U.P. 1987. Reaction kinetic parameters for ion transport from steady-state currentvoltage curves.Biophys. J. 51:569–585

    PubMed  Google Scholar 

  • Gray, M.A., Greenwell, J.R., Garton, A.J., Argent, B.E. 1990. Regulation of maxi-K+ channels on pancreatic duct cells by cyclic AMP-dependent phosphorylation.J. Membrane Biol. 115:203–215

    Article  Google Scholar 

  • Hagiwara, S., Miyazaki, S., Krasne, S., Ciani, S. 1977. Anomalous permeabilities of the egg cell membrane of a starfish in K+-Tl+ mixtures.J. Gen. Physiol. 70:269–281

    Article  PubMed  Google Scholar 

  • Hansen, U.P. 1990. Implications of control theory for homeostasis and phosphorylation of transport molecules.Bot. Acta 103:15–23

    Google Scholar 

  • Hansen, U.P., Dau, H., Vanselow K.H., Fisahn, J., Stein, S., Kolbowski, J. 1989. Thylakoid and plasmalemma fluxes.In: Plant Membrane Transport: The Current Position. J. Dainty, M.I. De Michelis, E. Marrè, and F. Rasi-Caldogno, editors. pp. 345–350. Elsevier, Amsterdam-New York-Oxford

    Google Scholar 

  • Hansen, U.-P., Fisahn, J. 1987.I-V-curve studies of the control of a K+-transporter inNitella by temperature.J. Membrane Biol. 98:1–13

    Article  Google Scholar 

  • Hansen, U.P., Gradmann, D., Sanders, D., Slayman, C.L. 1981. Interpretations of current-voltage relationships for “active” ion transport systems: I. Steady-state reaction-kinetic analysis of class-I mechanisms.J. Membrane Biol. 63:165–190

    Article  Google Scholar 

  • Hansen, U.-P., Tittor, J., Gradmann, D. 1983. Interpretation of current-voltage relationships for “active” ion transport systems: II. Nonsteady-state reaction-kinetic analysis of Class-I mechanisms with one slow time-constant.J. Membrane Biol. 75:141–169

    Google Scholar 

  • Hedrich, R., Schroeder, J.I. 1989. The physiology of ion channels and electrogenic pumps in higher plants.Ann. Rev. Plant Physiol. 40:539–569

    Article  Google Scholar 

  • Heinemann, S.H., Sigworth, F.J. 1990. Open channel noise: V. Fluctuating barriers to ion entry in gramicidin A channels.Biophys. J. 57:499–514

    PubMed  Google Scholar 

  • Hille, B., Schwarz, W. 1978. Potassium channels as multi-ion single-file pores.J. Gen. Physiol. 72:409–442

    PubMed  Google Scholar 

  • King, E.L., Altmann, C. 1956. A schematic method of deriving the rate laws for enzyme-catalyzed reactions.J. Phys. Chem. 60:1375–1378

    Article  Google Scholar 

  • Kolarev, J., Kulpa, J., Baijot, M., Goffeau, A. 1988. Characterization of a protein serine kinase from yeast plasma membrane.J. Biol. Chem. 263:10613–10619

    PubMed  Google Scholar 

  • Laver, D.R., Fairley, K.A., Walker, N.A. 1989. Ion permeation in a K+ channel inChara australis: Direct evidence for diffusion limitation of ion flow in a maxi-K channel.J. Membrane Biol. 108:153–164

    Article  Google Scholar 

  • Lühring, H. 1986. Recording of single K+ channels in the membrane of cytoplasmic drop ofChara australis.Protoplasma 133:19–28

    Article  Google Scholar 

  • McDonough, J.P., Mahler, H.P. 1982. Covalent phosphorylation of the Mg2+-dependent ATPase of yeast plasma membranes.J. Biol. Chem. 257:14579–14581

    PubMed  Google Scholar 

  • Negulyaev, Yu.A., Vedernikova, E.A., Savokhina, G.A. 1990. Aconitine-induced modification of single sodium channels in neuroblastoma cell membrane.Gen. Physiol. Biophys. 9:167–176

    PubMed  Google Scholar 

  • Nollau, V. 1979. Statistische Analysen. Birkhäuser, Basel-Stuttgart

    Google Scholar 

  • Pietrobon, D., Prod'hom, B., Hess, P. 1988. Conformational changes associated with ion permeation in L-type calcium channels.Nature 333:373–376

    PubMed  Google Scholar 

  • Portillo, F., Mazon, M.J. 1985. Activation of yeast plasma membrane ATPase by phorbol ester.FEBS Lett. 192:95–98

    Article  PubMed  Google Scholar 

  • Prod'hom, B., Pietrobon, D., Hess, P. 1987. Direct measurement of proton transfer rates to a group controlling the dihydropyridine-sensitive Ca2+ channel.Nature 239:243–246

    Article  Google Scholar 

  • Sachs, L. 1984. Applied Statistics. Springer-Verlag, New York-Berlin-Heidelberg-Tokyo

    Google Scholar 

  • Sakano, K., Tazawa, M. 1986. Tonoplast origin of the envelope membrane of cytoplasmic droplets prepared fromChara internodal cells.Protoplasma 131:247–249

    Article  Google Scholar 

  • Sorensen, H.W. 1980. Parameter Estimation. pp. 183–199. Marcel Dekker, New York-Basel

    Google Scholar 

  • Tester, M. 1988. Potassium channels in the plasmalemma ofChara corallina are multi-ion pores: Voltage-dependent blockade by Cs+ and anomalous permeabilities.J. Membrane Biol. 105:87–94

    Article  Google Scholar 

  • Tyerman, S.D., Findlay, G.P. 1989. Current-voltage curves of single Cl channels which coexist with two types of K channels in the tonoplast ofChara corallina.J. Exp. Bot. 40:105–117

    Google Scholar 

  • Vanselow, K.H., Hansen, U.P. 1989. The rapid effect of light on the K+ channel inNitella.J. Membrane Biol. 110:175–187

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Draber, S., Schultze, R. & Hansen, UP. Patch-clamp studies on the anomalous mole fraction effect of the K+ channel in cytoplasmic droplets ofNitella: An attempt to distinguish between a multi-ion single-file pore and an enzyme kinetic model with lazy state. J. Membrain Biol. 123, 183–190 (1991). https://doi.org/10.1007/BF01998088

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01998088

Key Words

Navigation