During oxidative phosphorylation most of the protons pumped out to the cytosol across the mitochondrial inner membrane return to the matrix through the ATP synthase, driving ATP synthesis. However, some of them leak back to the matrix through a proton-conductance pathway in the membrane. When the ATP synthase is inhibited with oligomycin and ATP is not being synthesized, all of the respiration is used to drive the proton leak. We report here that Mg2+ inhibits the proton conductance in rat skeletal-muscle mitochondria. Addition of Mg2+ inhibited both oligomycin-inhibited respiration and the proton conductance, while removal of Mg2+ using EDTA activated these processes. The proton conductance was inhibited by more than 80% as free Mg2+ was raised from 25 nM to 220 μM. Half-maximal inhibition occurred at about 1 μM free Mg2+, which is close to the contaminating free Mg2+ concentration in our incubations in the absence of added magnesium chelators. ATP, GTP, CTP, TTP or UTP at a concentration of 1 mM increased the oligomycin-inhibited respiration rate by about 50%. However, these NTP effects were abolished by addition of 2 mM Mg2+ and any NTP-stimulated proton conductance was explained completely by chelation of endogenous free Mg2+. The corresponding nucleoside diphosphates (ADP, GDP, CDP, TDP or UDP) at 1 mM had no effect on oligomycin-inhibited respiration. We conclude that proton conductance in rat skeletal-muscle mitochondria is very sensitive to free Mg2+ concentration but is insensitive to NTPs or NDPs at 1 mM.
Research Article| May 09 2000
Effects of magnesium and nucleotides on the proton conductance of rat skeletal-muscle mitochondria
Martin D. BRAND
Martin D. BRAND 1
*Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QW, U.K.
†MRC Dunn Human Nutrition Unit, Hills Road, Cambridge CB2 2XY, U.K.
1To whom correspondence should be addressed, at the MRC Dunn Human Nutrition Unit (e-mail martin.brand@;mrc-dunn.cam.ac.uk).
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Susana CADENAS, Martin D. BRAND; Effects of magnesium and nucleotides on the proton conductance of rat skeletal-muscle mitochondria. Biochem J 15 May 2000; 348 (1): 209–213. doi: https://doi.org/10.1042/bj3480209
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