Summary
Sudden respiratory blockade has been used to study rapid changes of the resting membrane potential, of intracellular adenosine 5′-triphosphate (ATP) levels, and of pyridine nucleotide reduction inNeurospora crassa. Membrane depolarization occurs with a first-order rate constant of 0.167 sec−1, following a lag period of about 4 sec, at 24°C (ambient temperature). This depolarization is several-fold too slow to be directly linked to electron transfer, as judged from the rate of pyridine nucleotide reduction, but has essentially the same rate constant as the decay of ATP. The latter process, however, shows no lag period after the respiratory inhibitor is introduced. Plots of membrane potential versus the intracellular ATP concentration yield saturation curves which are readily fitted by a Michaelis equation, to which is added a constant term representing the diffusion component of membrane potential. Parameters obtained from such fits indicate the maximal voltage which the pump can develop at high ATP levels to be 300 to 350 mV, with an apparentK 1/2 of 2.0mm. The data strongly suggest that an electrogenic ion pump in the plasma membrane ofNeurospora is fueled by ATP; comparison of the measured membrane potentials with the energy available from hydrolysis of ATP indicates that two ions could be pumped for each molecule of ATP split.
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References
Abrams, A. 1965. The release of bound adenosine triphosphatase from isolated bacterial membranes and the properties of the solubilized enzyme.J. Biol. Chem. 240:3675
Adrian, R. H. Slayman, C. L. 1966. Membrane potential and conductance during transport of sodium, potassium, and rubidium in frog muscle.J. Physiol. 184:970
Anderson, P. M., Wellner, V. P., Rosenthal, G. A., Meister, A. 1970. Carbonylphosphate synthetase (Escherichia coli).In: Methods in Enzymology. H. Tabor and C. W. Tabor, editors. Vol. XVII, p. 235: Metabolism of amino acids and amines, Part A. Academic Press, Inc., New York
Aubert, X., Chance, B., Keynes, R. D. 1964. Optical studies of biochemical events in the electric organ ofElectrophorus.Proc. Roy. Soc., B. 160:211
Brody, S. 1972. Regulation of pyridine nucleotide levels and ratios inNeurospora crassa.J. Biol. Chem. 247:6013
Chance, B., Williams, G. R. 1955. Respiratory enzymes in oxidative phosphorylation. IV. The respiratory chain.J. Biol. Chem. 217:429
Falcoz-Kelly, F., Cohen, G. N. 1970. Aspartokinase II and homoserine dehydrogenase II.In: Methods in Enzymology. H. Tabor and C. W. Tabor, editors. Vol. XVII, p. 699. Metabolism of amino acids and amines, Part A. Academic Press Inc., New York
Fein, H. 1966. Passing current through recording glass micropipette electrodes.IEEE Trans. Bio-med. Eng. 13:211
Fox, C. F. 1972. Membrane assembly.In: Membrane Molecular Biology. C. F. Fox and A. Keith, editors. p. 345. Sinaur Associates, Stamford, Connecticut
Gibson, Q. H. 1963. Inhibitors of gas transport.In: Metabolic Inhibitors: A Comprehensive Treatise. R. M. Hochester and J. H. Quastel, editors. Vol. II, p. 539. Academic Press, Inc., New York
Gonzalez, C. F., Shamoo, Y. E., Brodsky, W. A. 1967. Electrical nature of active chloride transport across short-circuited turtle bladders.Amer. J. Physiol. 212:641
Gradmann, D. 1970. Einfluß von Licht, Temperature and Außenmedium auf das elektrische Verhalten vonAcetabularia crenulata.Planta 93:323
Harold, F. M. 1962. Depletion and replenishment of the inorganic polyphosphate pools inNeurospora crassa.J. Bacteriol. 83:1047
Harold, F. M. 1972. Conservation and transformation of energy by bacterial membranes.Bact. Rev. 36:172
Hers, H. G. 1955. Fructokinase (ketokinase).In: Methods in Enzymology. S. P. Colowick and N. O. Kaplan, editors. Vol. I, p. 286. Academic Press Inc., New York
Hopfer, U., Lehninger, A. L., Thompson, T. E. 1968. Protonic conductance across lipid bilayer membranes induced by uncoupling agents for oxidative phosphorylation.Proc. Nat. Acad. Sci. 59:484
Kasbekar, D. K., Durbin, R. P. 1965. An adenosine triphosphatase from frog gastric mucosa.Biochim. Biophys. Acta 105:472
Kernan, R. P. 1970. Electrogenic or linked transport.In: Membranes and Ion Transport. E. E. Bittar, editor. Vol. I, p. 395. Wiley-Interscience, New York
Kitasato, H. 1968. The influence of H+ on the membrane potential and ion fluxes ofNitella.J. Gen. Physiol. 52:60
Lambowitz, A. M., Slayman, C. W., Slayman, C. L., Bonner, W. D. 1972. The electron transport components of wild type and Poky strains ofNeurospora crassa.J. Biol. Chem. 247:1536
Lester, G., Stone, D., Hechter, O. 1958. The effects of deoxycorticosterone and other steroids onNeurospora crassa.Arch. Biochem. Biophys. 75:196
MacRobbie, E. A. C. 1970. The active transport of ions in plant cells.Quart. Rev. Biophys. 3:251
Marmor, M. F. 1971. The independence of electrogenic sodium transport and membrane potential in a molluscan neurone.J. Physiol. 218:599
Marquardt, D. W. 1963. An algorithm for least-squares estimation on non-linear parameters.J. Soc. Ind. Appl. Math. 11:431
Marsh, C., Militzer, W. 1956. Thermal enzymes. VII: Further data on an adenosine triphosphatase.Arch. Biochem. Biophys. 60:433
Mitchell, P. 1966. Chemiosmotic coupling in oxidative and photosynthetic phosphorylation.Biol. Rev. 41:445
Mitchell, P. 1970. Reversible coupling between transport and chemical reactions.In: Membranes and Ion Transport. E. E. Bittar, editor. Vol. I, p. 192. Wiley-Interscience, New York
Phillips, R. C., George, P., Rutman, R. J. 1969. Thermodynamic data for the hydrolysis of adenosine triphosphate as a function of pH, Mg2+ ion concentration, and ionic strength.J. Biol. Chem. 241:3330
Raven, J. A. 1968. Action spectra for photosynthesis and light-stimulated ion transport processes inHydrodictyon africanum.New Phytol. 68:45
Raven, J. A. 1968. Effects of inhibitors on photosynthesis and the active influxes of K and Cl inHydrodictyon africanum.New Phytol. 68:1089
Rehm, W. S. 1966. Electrogenic mechanism of the frog's gastric mucosa.Ann. N.Y. Acad. Sci. 137:591
Rehm, W. S. 1972. Proton transport.In: Metabolic Pathways. VI. Metabolic Transport. L. E. Hokin, editor. p. 187. Academic Press Inc., New York
Ritchie, J. M. 1971. Electrogenic ion pumping in nervous tissue.Curr. Top. Bioenerget. 4:327
Roon, R. J., Levenberg, B. 1970. ATP: Urea amidolyase (ADP) (Candida utilis).In: Methods in Enzymology. H. Tabor and C. W. Tabor, editors. Vol. XVII, p. 317. Metabolism of amino acids and amines, Part A. Academic Press Inc., New York
Saddler, H. D. W. 1970. The membrane potential ofAcetabularia mediterranea.J. Gen. Physiol. 55:802
Schoner, W., Beusch, R., Kramer, R. 1968. On the mechanism of Na+- and K+-stimulated hydrolysis of adenosine triphosphate. 2. Comparison of nucleotide specificities of Na+- and K+-activated ATPase and Na+-dependent phosphorylation of cell membranes.Europ. J. Biochem. 7:102
Skulachev, V. P. 1971. Energy transformations in the respiratory chain.Curr. Top. Bioenerget. 4:127
Slayman, C. L. 1965. Electrical properties ofNeurospora crassa: Effects of external cations on the intracellular potential.J. Gen. Physiol. 49:69
Slayman, C. L. 1965. Electrical properties ofNeurospora crassa: Respiration and the intracellular potential.J. Gen. Physiol. 49:93
Slayman, C. L. 1970. Movement of ions and electrogenesis in microorganism.Amer. Zoologist 10:377
Slayman, C. L. 1973. Adenine nucleotide levels inNeurospora, as influenced by conditions of growth and by metabolic inhibitors.J. Bacteriol. 114:752
Slayman, C. L., Lu, C.Y.-H., Shane, L. 1970. Correlated changes in membrane potential and ATP concentrations inNeurospora.Nature 226:274
Slayman, C. L., Slayman, C. W. 1968. Net uptake of potassium inNeurospora: Exchange for sodium and hydrogen ions.J. Gen. Physiol. 52:424
Slayman, C. L., Slayman, C. W. 1973. H+-dependent cotransport and the electrogenic pump in the plasma membrane ofNeurospora. Abstracts,73 rd Ann. Meet. Amer. Soc. Microbiol., Item P189
Slayman, C. W., Slayman, C. L. 1970. Potassium transport inNeurospora: Evidence for a multisite carrier at high pH.J. Gen. Physiol. 55:758
Slayman, C. W., Tatum, E. L. 1964. Potassium transport inNeurospora. I. Intracellular sodium and potassium concentrations, and cation requirements for growth.Biochim. Biophys. Acta 88:578
Slayman, C. W., Tatum, E. L. 1965. Potassium transport inNeurospora. II. Measurement of steady-state potassium fluxes.Biochim. Biophys. Acta 102:149
Spanswick, R. M. 1972. Evidence for an electrogenic ion pump inNitella translucens. I. The effects of pH, K+, Na+, light, and temperature on the membrane potential and resistance.Biochim. Biophys. Acta 288:73
Strehler, B. L. 1965. Adenosine-5′-triphosphate and creatine phosphate. Determination with luciferase.In: Methods of Enzymatic Analysis. H. U. Bergemeyer, editor. p. 559. Academic Press Inc., New York
Tamai, T., Kagiyama, S. 1968. Studies of cat heart muscle during recovery after prolonged hypothermia. Hyperpolarization of cell membranes and its dependence on the sodium pump with electrogenic characteristics.Circulation Res. 22:423
Thomas, R. C. 1969. Membrane current and intracellular sodium changes in a snail neurone during extrusion of injected sodium.J. Physiol. 201:495
Thomas, R. C. 1972. Electrogenic sodium pump in nerve and muscle cells.Physiol. Rev. 52:563
Ullrich, F. 1966. The inhibition of magnesium-activated mitochondrial adenosine triphosphatase by calcium.Biochim. Biophys. Acta 122:298
Viotti, A., Bagni, N., Sturani, E., Alberghina, F. A. M. 1971. Magnesium and polyamine levels inNeurospora crassa mycelia.Biochim. Biophys. Acta 244:329
Vogel, H. J. 1956. A convenient growth medium forNeurospora (Medium N).Microbiol. Gen. Bull. 13:42
Weibull, C., Greenawalt, J. W., Löw, H. 1962. The hydrolysis of adenosine triphosphate by all fractions ofBacillus megaterium. I. Localization and general characteristics of the enzymatic activities.J. Biol. Chem. 237:847
West, I. C., Mitchell, P. 1973. Stoichiometry of lactose-proton symport across the plasma membrane ofEscherichia coli.Biochem. J. 132:587
Zalokar, M. 1959. Enzyme activity and cell differentiation inNeurospora.Amer. J. Bot. 46:555
Zalokar, M. 1959. Growth and differentiation ofNeurospora hyphae.Amer. J. Bot. 46:602
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Slayman, C.L., Long, W.S. & Lu, C.Y.H. The relationship between ATP and an electrogenic pump in the plasma membrane ofNeurospora crassa . J. Membrain Biol. 14, 305–338 (1973). https://doi.org/10.1007/BF01868083
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DOI: https://doi.org/10.1007/BF01868083