Altered inhibition of the rat skeletal ryanodine receptor/calcium release channel by magnesium in the presence of ATP.
Magnesium-induced inhibition of the skeletal ryanodine receptor/calcium-release channel (RyR) was studied in the presence and absence of ATP under isolated conditions and in situ, by examining the RyR incorporated into a planar lipid bilayer and the calcium release flux (Rrel) in isolated single fibres mounted in the double Vaseline gap system. When the incorporated RyR had been activated by calcium (50 microM) in the absence of ATP, the magnesium-induced inhibition showed co-operativity with a Hill coefficient (N) of 1.83 and a half-inhibitory concentration (IC50) of 635 microM. When the open probability was measured in the presence of 5 mM ATP and at a low calcium concentration, the magnesium-induced inhibition was non-cooperative (N=1.1, IC50= 860 microM). In isolated muscle fibres, in the presence of ATP, lowering the intracellular magnesium concentration ([Mg2+]i) increased the maximal Rrel and shifted its voltage dependence to more negative membrane potentials. Increasing [Mg2+]i had the opposite effect. The concentration dependence was described with an IC50 of 174 microM, N=1, under depolarized conditions and showed a tenfold increase in affinity in polarized fibres. At the concentration required for the measurements from isolated fibres, ATP had a full activatory effect on the isolated channel. At a low calcium concentration, the RyR had two ATP-binding sites with half-activatory concentrations of 19 and 350 microM, respectively.
Jóna
I
I
Department of Physiology, University of Debrecen, Medical School, Hungary. jonai@phys.dote.hu
Szegedi
C
C
Sárközi
S
S
Szentesi
P
P
Csernoch
L
L
Kovács
L
L
D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D066298:In Vitro Techniques; D015640:Ion Channel Gating / Q000187:drug effects* / Q000502:physiology; D008274:Magnesium / Q000494:pharmacology*; D018485:Muscle Fibers, Skeletal / Q000502:physiology; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D019837:Ryanodine Receptor Calcium Release Channel / Q000378:metabolism*; D012519:Sarcoplasmic Reticulum / Q000378:metabolism
D019837:Ryanodine Receptor Calcium Release Channel; D000255:Adenosine Triphosphate; D008274:Magnesium