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  • 1
    ISSN: 1432-2013
    Keywords: Calcium channel blockers ; Cytosolic calcium ; Sodium pump ; Muscle fibres
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The effects of 10 mM Ca2+ and Ca2+ channel blockers verapamil, diltiazem and flunarizine on the ouabain-sensitive electrogenic Na+, K+ pump activity of mouse diaphragm muscle fibres enriched with Na+ were compared with the changes in cytosolic [Ca2+]. The electrogenic Na+ pump activity produced by adding K+ to muscles previously bathed for 4 h in a K+-free, 2-mM [Ca2+] solution increased the resting membrane potential by about 18 mV. This hyperpolarization was completely inhibited after 10 min incubation in 10 mM Ca2+. Verapamil 10−5M, 10−5M diltiazem and 10−7 M flunarizine effectively prevented the effect of elevated [Ca2+]. At these concentrations, these drugs did not affect the K+-induced hyperpolarization. In mouse diaphragm, the basal cytosolic [Ca2+] measured by the fluorescent indicator 1-[2-(5-carboxyoxazol-2-yl)-6-aminobenzofuran-5-oxy]2-(2′-amino 5′-methylphenoxy) ethane-N,N,N′,N′-tetraacetic acid acetoxymethyl ester (fura-2/AM) was 261±6 nM. After 4 h in a Liley K+-free, 2 mM [Ca2+] solution, the cytosolic [Ca2+] increased to 314±28 nM. Increase in [Ca2+] from 2 to 10 mM caused a twofold increase of cytosolic [Ca2+] to 637±26 nM. This rise was, like the Ca2+-induced inhibition of electrogenic pump, prevented by 10−5 M verapamil, 10−5M diltiazem and 10−7 M flunarizine. The results suggest that substances which block Ca2+ entry into the cell prevent the Ca2+ induced inhibition of the Na+ pump.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-4919
    Keywords: (Na/K)-ATPase ; electrophilic reagents ; cation binding site ; isolated perfused heart ; 5-nitrofurylethylene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The effect of electrophilic substances: p-bromophenylisothiocyanate (PBITC); fluoresceinisothiocyanate (FITC); [4-isothiocyanatophenyl-(6-thioureidohexyl)-carbamoylmethyl]-ATP (ATPITC); 2,4,6-trinitrobezenesulfonic acid (TNBS); 1-(5-nitro-2-furyl)-2-phenylsulfonyl-2-furylcarbonyl ethylene (FE1); 1-(5-phenylsulfonyl-2-furyl)-2-phenylsulfonyl-2-furylcarbonyl ethylene (FE2) and 1-(5-phenylsulfonyl-2-furyl)-2-phenylsulfonyl-2-tienocarbonyl ethylene (FE3) on the sarcolemmal (Na/K)-ATPase isolated from guinea-pig hearts was studied. FITC and PBITC were found to inhibit competitively the activation of (Na/K)-ATPase by ATP. Being for the enzyme inhibitor and substrate at the same time ATPITC does not offered clear kinetic behavior. However, the activation of (Na/K)-ATPase by sodium and potassium ions was inhibited non-competitively by all three isothiocyanates. These data indicated that isothiocyanates may interact predominantly in the ATP-binding site of the enzyme molecule. In contrary to isothiocyanates TNBS and FE1 (FE2 and FE3 were ineffective) inhibited the activation of (Na/K)-ATPase by ATP non-competitively i.e., their interaction in the ATP-binding site seemed to be improbable. Nevertheless, TNBS and FE1 both manifested affinities to that moiety of (Na/K)-ATPase molecule which is binding potassium. More specific was the effect of FE1 that showed clearly competitive inhibition of potassium-stimulation of the enzyme activity. FE1 exerted also an ouabain-like effect on the mechanical activity of isolated perfused guinea-pig heart. This result indicates that FE1 seems to exert a selective inhibition of the (Na/K)-ATPase not only in vitro but also in integrated cardiac tissue.
    Type of Medium: Electronic Resource
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