Effects of pH changes on sodium pump fluxes in squid giant axon

1987 ◽  
Vol 253 (4) ◽  
pp. C547-C554 ◽  
Author(s):  
G. E. Breitwieser ◽  
A. A. Altamirano ◽  
J. M. Russell
Keyword(s):  
Sodium Pump ◽  
Maximum Velocity ◽  
Activation Parameters ◽  
Giant Axon ◽  
Inhibitory Effect ◽  
Squid Giant Axon ◽  
Extracellular Ph ◽  
Sodium Efflux ◽  
Effects Of Ph ◽  
Ph Range

The effects of independently varying intracellular and extracellular pH on sodium pump fluxes were studied in the squid giant axon. By means of intracellular dialysis, we found that changes of intracellular pH (pHi), but not of extracellular pH, affected ouabain-sensitive Na+ efflux and K+ influx over the pH range of 6.0-8.6. Both fluxes were maximum at a pHi of 7.2-7.4. Variations away from this optimal pHi in either the acidic or alkaline direction resulted in a graded inhibition of both ouabain-sensitive fluxes. The kinetic basis for the inhibitory effect of acidic pHi was examined by comparing the kinetic parameters of activation of ouabain-sensitive sodium efflux by intracellular Na+ (Na+i) and extracellular K+ (K+o) at normal pHi with those at acidic pHi. We found that the inhibitory effect of intracellular acidity results from a reversible decrease in maximum velocity (Vmax), without an effect on the activation parameters for Na+i (K1/2 Na+i) or K+o (K1/2 K+o).

Electric current generated by squid giant axon sodium pump: external K and internal ADP effects

1978 ◽  
Vol 235 (1) ◽  
pp. C63-C68 ◽  
Author(s):  
R. F. Abercrombie ◽  
P. de Weer
Keyword(s):  
Sodium Pump ◽  
Giant Axon ◽  
Pump Current ◽  
Membrane Resistance ◽  
Extracellular Potassium ◽  
Sodium Efflux ◽  
Steroid Sensitive ◽  
External Potassium ◽  
Loligo Pealei ◽  
External K

The operation of the sodium pump of giant axons of the squid, Loligo pealei, has been studied simultaneously in two independent ways: 1) by measuring sodium efflux with 22Na, and 2) by calculating the transmembrane current generated by the pump from measurements of membrane resistance and digitalis-sensitive membrane potential. In normal, untreated axons, the effect of increasing the external potassium concentration on both sodium efflux and pump current is similar, which suggests that Na:K pump stoichiometry remains relatively constant in the range of 0-20 mM external K. The data are compatible with a 3:2 Na:K ratio. In axons whose intracellular ADP level has been elevated by injection of L-arginine, a large, electrically silent, cardiotonic steroid-sensitive sodium efflux takes place in the absence of external potassium; this suggests that pump-mediated Na:Na exchange is 1:1 or electroneutral. Finally, elevation of external potassium levels causes the appearance, in high-ADP axons, of electrogenic pumping, with little effect on sodium efflux; hence, in contrast to what is seen in normal (low-ADP) axons, the charge translocated, per sodium ion extruded, increases sharply with increasing extracellular potassium levels.

scholarly journals Strophanthidin-sensitive sodium fluxes in metabolically poisoned frog skeletal muscle.

1976 ◽  
Vol 68 (4) ◽  
pp. 405-420 ◽  
Author(s):  
B G Kennedy ◽  
P De Weer
Keyword(s):  
Skeletal Muscle ◽  
Membrane Potential ◽  
Human Erythrocyte ◽  
Sodium Pump ◽  
Giant Axon ◽  
Squid Giant Axon ◽  
Frog Skeletal Muscle ◽  
Na Efflux ◽  
Unidirectional Fluxes ◽  
External K

Strophanthidin-sensitive and insensitive unidirectional fluxes of Na were measured in fog sartorius muscles whose internal Na levels were elevated by overnight storage in the cold. ATP levels were lowered, and ADP levels raised, by metabolic poisoning with either 2,4-dinitrofluorobenzene or iodoacetamide. Strophanthidin-sensitive Na efflux and influx both increased after poisoning, while strophanthidin-insensitives fluxes did not. The increase in efflux did not require the presence of external K but was greatly attenuated when Li replaced Na as the major external cation. Membrane potential was not markedly altered by 2,4-dinitrofluorobenzene. These observations indicate that the sodium pump of frog skeletal muscle resembles that of squid giant axon and human erythrocyte in its ability to catalyze Na-Na exchange to an extent determined by intracellular ATP/ADP levels.

scholarly journals Effects of Intracellular Adenosine-5'-diphosphate and Orthophosphate on the Sensitivity of Sodium Efflux from Squid Axon to External Sodium and Potassium

1970 ◽  
Vol 56 (5) ◽  
pp. 583-620 ◽  
Author(s):  
Paul De Weer
Keyword(s):  
Creatine Phosphokinase ◽  
Giant Axon ◽  
Squid Giant Axon ◽  
Squid Axon ◽  
Sodium Efflux ◽  
Excess Mg ◽  
Sodium And Potassium ◽  
External K

A study was made of sodium efflux from squid giant axon, and its sensitivity to external K and Na. When sodium efflux from untreated axons was strongly stimulated by Ko, Nao was inhibitory; when dependence on Ko was low, Nao had a stimulatory effect. Incipient CN poisoning or apyrase injection, which produces high intracellular levels of ADP1 and Pi, rendered sodium efflux less dependent on external K and more dependent on external Na. Injection of ADP, AMP, arginine, or creatine + creatine phosphokinase, all of which raise ADP levels without raising Pi levels, had the same effect as incipient CN poisoning. Pi injection had no effect on the K sensitivity of sodium efflux. Axons depleted of arginine and phosphoarginine by injection of arginase still lost their K sensitivity when the ATP:ADP ratio was lowered and regained it partially when the ratio was raised. Rough calculations show that sodium efflux is maximally Ko-dependent when the ATP:ADP ratio is about 10:1, becomes insensitive to Ko when the ratio is about 1:2, and is inhibited by Ko when the ratio is about 1:10. Deoxy-ATP mimicked ADP when injected into intact axons. Excess Mg, as well as Pi, inhibited both strophanthidin-sensitive and strophanthidin-insensitive sodium efflux. An outline is presented for a model which might explain the effects of ADP, Pi and deoxy-ATP.

Effects of Intracellular ADP and Pi on the Sodium Pump of Squid Giant Axon

Nature ◽  
1970 ◽  
Vol 226 (5252) ◽  
pp. 1251-1252 ◽  
Author(s):  
P. DE WEER
Keyword(s):  
Sodium Pump ◽  
Giant Axon ◽  
Squid Giant Axon

Electrogenic Sodium Pump in Squid Giant Axon

Science ◽  
1973 ◽  
Vol 179 (4080) ◽  
pp. 1326-1328 ◽  
Author(s):  
P. De Weer ◽  
D. Geduldig
Keyword(s):  
Sodium Pump ◽  
Giant Axon ◽  
Squid Giant Axon ◽  
Electrogenic Sodium Pump

scholarly journals Axoplasmic free magnesium levels and magnesium extrusion from squid giant axons.

1976 ◽  
Vol 68 (2) ◽  
pp. 159-178 ◽  
Author(s):  
P De Weer
Keyword(s):  
Sodium Pump ◽  
Giant Axon ◽  
Magnesium Content ◽  
Magnesium Concentration ◽  
Citrate Buffer ◽  
Sodium Efflux ◽  
Loligo Pealei ◽  
Free Magnesium ◽  
Total Magnesium ◽  
Very High

The free magnesium concentration in the axoplasm of the giant axon of the squid, Loligo pealei, was estimated by exploting the known sensitivity of the sodium pump to intracellular Mg2+ levels. The Mg-citrate buffer which, when injected into the axon, resulted in no change in sodium efflux was in equilibrium with a Mg2+ level of about 3--4 mM. Optimal [Mg2+] for the sodium pump is somewhat higher. Total magnesium content of axoplasm was 6.7 mmol/kg, and that of hemolymph was 44 mM. The rate coefficient for 28Mg efflux was about 2 X 10(-3) min-u for a 500-mum axon at 22-25degreesC, with a very high temperature coefficient (Q10=4-5). This efflux is inhibited 95% by injection of apyrase and 75% by removal of external sodium, and seems unaffected by membrane potential or potassium ions. Increased intracellular ADP levels do not affect Mg efflux nor its requirement for Na+/o, but extracellularl magnesium ions do. Activation of 28Mg efflux by Na+/o follows hyperbolic kinetics, with Mg2+/o reducing the affinity of the system for Na+/o. Lanthanum and D600 reversibly inhibit Mg efflux. In the absence of both Na+ and Mg2+, but not in their presence, removal of Ca2+ from the seawater vastly increased 28Mg efflux; this efflux was also strongly inhibited by lanthanum. A small (10(-14) mol cm-2) extra Mg efflux accompanies the conduction of an action potential.

scholarly journals Sodium efflux in Myxicola giant axons.

1977 ◽  
Vol 69 (6) ◽  
pp. 765-778 ◽  
Author(s):  
R F Abercrombie ◽  
R A Sjodin
Keyword(s):  
Membrane Transport ◽  
Giant Axon ◽  
Squid Giant Axon ◽  
Sodium Efflux ◽  
Giant Axons ◽  
Na Pump ◽  
Transport Studies ◽  
All Solutions ◽  
Na Efflux

Several properties of the Na pump in giant axons from the marine annelid Myxicola infundibulum have been determined in an attempt to characterize this preparation for membrane transport studies. Both NaO and KO activated the Na pump of normal microinjected Myxicola axons. In this preparation, the KO activation was less and the NaO activation much greater than that found in the squid giant axon. However, when the intracellular ATP:ADP ratio of the Myxicola axon was elevated by injection of an extraneous phosphagen system, the K sensitivity of Na efflux increased to the magnitude characteristic of squid axons and the activating effect of NaO disappeared. Several axons were injected with Na2SO4 in order to determine the effect of elevated Nai on the Na efflux. Increasing Nai enhanced a component of Na efflux which was insensitive to ouabain and dependent on [Ca] in Na-free (Li) seawater. After subtracting the CaO-dependent fraction, Na efflux was related linearly to [Na]i in all solutions except in K-free (Li) seawater, where it appeared to reach saturation at high [Na]i.

scholarly journals Acyl Coenzyme A Synthetase from Pseudomonas fragi Catalyzes the Synthesis of Adenosine 5′-Polyphosphates and Dinucleoside Polyphosphates

1998 ◽  
Vol 180 (12) ◽  
pp. 3152-3158 ◽  
Author(s):  
Rui Fontes ◽  
Maria A. Günther Sillero ◽  
Antonio Sillero
Keyword(s):  
Coenzyme A ◽  
Divalent Cations ◽  
Maximum Velocity ◽  
Inhibitory Effect ◽  
Maximum Activity ◽  
Pseudomonas Fragi ◽  
Ph Values ◽  
Ph Range ◽  
Acyl Coenzyme A ◽  
Dinucleoside Polyphosphates

ABSTRACT Acyl coenzyme A (CoA) synthetase (EC 6.2.1.8 ) fromPseudomonas fragi catalyzes the synthesis of adenosine 5′-tetraphosphate (p4A) and adenosine 5′-pentaphosphate (p5A) from ATP and tri- or tetrapolyphosphate, respectively. dATP, adenosine-5′-O-[γ-thiotriphosphate] (ATPγS), adenosine(5′)tetraphospho(5′)adenosine (Ap4A), and adenosine(5′)pentaphospho(5′)adenosine (Ap5A) are also substrates of the reaction yielding p4(d)A in the presence of tripolyphosphate (P3). UTP, CTP, and AMP are not substrates of the reaction. The Km values for ATP and P3 are 0.015 and 1.3 mM, respectively. Maximum velocity was obtained in the presence of MgCl2 or CoCl2 equimolecular with the sum of ATP and P3. The relative rates of synthesis of p4A with divalent cations were Mg = Co > Mn = Zn >> Ca. In the pH range used, maximum and minimum activities were measured at pH values of 5.5 and 8.2, respectively; the opposite was observed for the synthesis of palmitoyl-CoA, with maximum activity in the alkaline range. The relative rates of synthesis of palmitoyl-CoA and p4A are around 10 (at pH 5.5) and around 200 (at pH 8.2). The synthesis of p4A is inhibited by CoA, and the inhibitory effect of CoA can be counteracted by fatty acids. To a lesser extent, the enzyme catalyzes the synthesis also of Ap4A (from ATP), Ap5A (from p4A), and adenosine(5′)tetraphospho(5′)nucleoside (Ap4N) from adequate adenylyl donors (ATP, ATPγS, or octanoyl-AMP) and adequate adenylyl acceptors (nucleoside triphosphates).

Sign in / Sign up

Export Citation Format

Share Document