Inhibition of (Na+ + K+)-ATPase by ouabain: Involvement of calcium and membrane proteins

Lelièvre, L.; Zachowski, Alain; Charlemagne, D.; Laget, P; Paraf, A

doi:10.1016/0005-2736(79)90338-9

Cited by 37 publications

(15 citation statements)

References 18 publications

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“…The biphasic dose response curve observed at 1 pM free (Ca2+) (Fig. 3) did not correspond to that previously reported with EDTA-treated membranes [9]. The use of an EGTACa2+ buffer in the data presented here could account for the difference observed, since in our previous work [9], we referred to added Caz+ and not free Ca2'.…”

Section: Specific Involvement Of Calmodulincontrasting

confidence: 67%

“…(0 They bound on domains distinct from the Na',K '-ATPase since the amount of Tm or &actinin necessary for a complete shift was independent of the specific activity of the membrane preparations: 0.33 & 0.03 nmol/mg of membrane fractions, the activities of which varied from 20 in EDTAtreated control membranes [7,9,101 to 42 pmol P i x h-l x mg-' (Table 1). Assuming a molecular mass of 68 kDa and 280 kDa, respectively for /$actinin (or Tm) and Na+,K+-ATPase, the /?-actinin (or Tm) and Na+,K '-ATPase molar ratios varied from 10 to 4.…”

Section: Similurity Between Effects Of Tm and P-actininmentioning

confidence: 99%

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Specific involvement of calmodulin and non‐specific effect of tropomyosin in the sensitivity to ouabain of Na⁺, K⁺‐ATPase in murine plasmocytoma cells

Lelièvre

Potter

Piascik

et al. 1985

European Journal of Biochemistry

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The Kd for ouabain for inhibition of Na+, K+‐ATPase isolated from murine plasmocytoma MOPC 173 cells is 120 μM, but when isolated in the presence of EDTA, it is 100‐fold lower (1.2 μM). Simultaneous addition of muscle tropomyosin and calcium to sensitive membranes restored the original insensitivity (tropomyosin bound to the membranes in an irreversible and saturable manner). For comparison 86Rb influx into intact cells, mediated by the Na+, K+‐pump, is half‐maximally inhibited at 50 μM ouabain. Calcium converts the enzyme to an insensitive form. This appeared to involve calmodulin because after extraction of calmodulin with EDTA and EGTA from sensitive membranes, they could not be made insensitive by the addition of tropomyosin and Ca2+. Addition of exogenous calmodulin to these calmodulin‐depleted membranes was required, in addition to tropomyosin and Ca2+, to decrease the ouabain sensitivity. The involvement of calmodulin was further assessed by measuring the range of Ca2+ concentrations required to convert to the insensitive form. At saturating concentrations of tropomyosin, increasing free [Ca2+] up to 3 μM led to an heterogenous population of Na+, K+‐ATPase forms. The calcium dependency was a saturable process. The shift to the insensitive form was half maximal at 0.65 + 0.11 μM free Ca2+ and was abolished by the addition of troponin I or trifluoroperazine (0.1 mM). These results suggest that, in murine plasmocytoma cells, the intrinsic sensitivity of Na+, K+‐ATPase to ouabain might be regulated by a calmodulin‐dependent process within a submembrane contractile‐like environment.

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Section: Specific Involvement Of Calmodulincontrasting

confidence: 67%

Section: Similurity Between Effects Of Tm and P-actininmentioning

confidence: 99%

Specific involvement of calmodulin and non‐specific effect of tropomyosin in the sensitivity to ouabain of Na⁺, K⁺‐ATPase in murine plasmocytoma cells

Lelièvre

Potter

Piascik

et al. 1985

European Journal of Biochemistry

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“…In cells from rodents and other digitalis-resistant species, cardiac glycosides reach equilibrium in about 5 min (Lelievre et al, 1979;Mansier & Lelievre, 1982). A flux experiment of 10-15 min (see Methods) cannot reveal the high association rate constants of genins in rat erythrocytes or mouse macrophages.…”

Section: Discussionmentioning

confidence: 98%

High sensitivity of the Na⁺, K⁺‐pump of human red blood cells to genins of cardiac glycosides

Senn

Lelièvre

Braquet³

et al. 1988

British J Pharmacology

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1 Four different cardiac glycosides (ouabain, digitoxin, digoxin and gitoxin) and their corresponding genins were tested on Na+, K+ -pump fluxes measured under steady-state and initial rate conditions (non equilibrium conditions) in human and rat erythrocytes and in mouse macrophages.2 In human red cells, Na+, K+-pump fluxes exhibited up to 8 fold higher sensitivity to genins than to glycosides. In addition genins, but not the corresponding glycosides, exhibited double reactivity with regard to the erythrocyte Na+, K+-pump (with the exception of gitoxigenin). A weak reactivity component was similar to the one of the corresponding glycosides (IC5O of about 10-6 M) and a high reactivity component exhibited IC5o values varying from 0.1 to 0.5 x 106 M for digitoxigenin and ouabagenin respectively.3 In contrast with human red cells, the initial rate of Na+, K+-pump fluxes in rat erythrocytes and mouse macrophages was less sensitive to genins than to the corresponding cardiac glycosides. 4 Dihydroouabain was 3, 10 and 75 times less active than ouabain in inhibiting the initial rate of Na+, K +-pump fluxes in human and rat erythrocytes and in mouse macrophages respectively. 5 In conclusion, Na+, K+-pump fluxes measured under initial rate conditions in human erythrocytes exhibit an unusually high sensitivity to genins of cardiac glycosides. This property probably results from the fast binding rate constants of genins and the slow association rates of glycosides to human red cells.

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“…More recently, Lelievre, Paraf, Charlemagne & Sheppard (1977) using wild type and ouabain-resistant plasmocytoma cell lines, suggested that adenylate cyclase and Na+, ©) Macmillan Publishers Ltd 1980 K+-ATPase may be closely coordinated in control of membrane activity and that a protein of Mr 37,000 may control the action of ouabain on the Na+, K +-ATPase with respect to digitalis resistance (Lelievre, Zachowski, Laget, Charlemagne & Paraf, 1979). Merrell Research Center has prepared a new compound cis-N-(2-phenylcyclopentyl) azacyclotridecl-en-2-amine monohydrochloride coded RMI 12330A, which has been reported to inhibit hormonestimulated and basal adenylate cyclase of rat liver plasma membrane preparations (Siegel & Wiech, 1977;Guellaen, Mahu, Mavier, Berthelot & Hanoune, 1977).…”

Section: Introductionmentioning

confidence: 99%

Effects of Rmi 12330a, a New Inhibitor of Adenylate Cyclase on Myocardial Function and Subcellular Activity

Grupp

Johnson

et al. 1980

British J Pharmacology

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RMI 12330A, a lactam‐imine, at concentrations of 10−4 m and higher, inhibited basal as well as isoprenaline and NaF‐stimulated adenylate cyclase activity of guinea‐pig heart homogenates. However, RMI 12330A was a more potent inhibitor of histamine‐stimulated adenylate cyclase (IC50 of 1.5 × 10−5 m). In the isolated work‐performing heart of the guinea‐pig, RMI 12330A (IC50 of 1.1 × 10−6 m) depressed all cardiac functions: pressures developed, dP/dt, contractile force, dF/dt, work performance and stroke work. Left atrial pressure rose and the positive inotropic response to increasing heart rate (staircase) became negative. Histamine, isoprenaline and ouabain no longer caused positive inotropic effects. Increasing the perfusate calcium concentration from 2.5 mm to 4.5 and 6.5 mm completely restored cardiac function after its depression by RMI 12330A. RMI 12330A uncoupled mitochondrial oxidative phosphorylation; the classical uncoupler, dinitrophenol, had the same effects on cardiac dynamics as RMI 12330A. RMI in high doses inhibited hydrolytic activity of Na+, K+‐ATPase of crude and purified heart preparations (IC50 of 1.7 × 10−4 m) and inhibited ouabain binding to the same enzymes (IC50 of 1.5 × 10−4 m). A lactam‐imine analogue of RMI 12330A that had no effect on adenylate cyclase, was also without effect on any of the systems examined.

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Inhibition of (Na+ + K+)-ATPase by ouabain: Involvement of calcium and membrane proteins

Cited by 37 publications

References 18 publications

Specific involvement of calmodulin and non‐specific effect of tropomyosin in the sensitivity to ouabain of Na⁺, K⁺‐ATPase in murine plasmocytoma cells

Specific involvement of calmodulin and non‐specific effect of tropomyosin in the sensitivity to ouabain of Na⁺, K⁺‐ATPase in murine plasmocytoma cells

High sensitivity of the Na⁺, K⁺‐pump of human red blood cells to genins of cardiac glycosides

Effects of Rmi 12330a, a New Inhibitor of Adenylate Cyclase on Myocardial Function and Subcellular Activity

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Inhibition of (Na+ + K+)-ATPase by ouabain: Involvement of calcium and membrane proteins

Cited by 37 publications

References 18 publications

Specific involvement of calmodulin and non‐specific effect of tropomyosin in the sensitivity to ouabain of Na+, K+‐ATPase in murine plasmocytoma cells

Specific involvement of calmodulin and non‐specific effect of tropomyosin in the sensitivity to ouabain of Na+, K+‐ATPase in murine plasmocytoma cells

High sensitivity of the Na+, K+‐pump of human red blood cells to genins of cardiac glycosides

Effects of Rmi 12330a, a New Inhibitor of Adenylate Cyclase on Myocardial Function and Subcellular Activity

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Specific involvement of calmodulin and non‐specific effect of tropomyosin in the sensitivity to ouabain of Na⁺, K⁺‐ATPase in murine plasmocytoma cells

Specific involvement of calmodulin and non‐specific effect of tropomyosin in the sensitivity to ouabain of Na⁺, K⁺‐ATPase in murine plasmocytoma cells

High sensitivity of the Na⁺, K⁺‐pump of human red blood cells to genins of cardiac glycosides