Effects of delipidation on proton translocation and ATPase activity in beef heart electron transport particles.
Delipidation of beef heart electron transport particles with phospholipase A2 has been examined. When the particles were treated with the lipase and subjected to a low bovine serum albumin wash, ATPase activity was unaffected as was the lipid/protein ratio of the particles. However, energisation by ATP/Mg2+ was abolished. Furthermore, unsaturated but not saturated fatty acids discharged the steady-state ATP-driven membrane potential of control samples. When the phospholipase A2 hydrolysis products were removed, inhibition of energy-linked reactions in the lipid-depleted particles was still observed and was interpreted in terms of non-specific leaks in the vesicle membranes, and 'specific' leaks through impaired H+-ATPase complexes. ATPase activity was less susceptible to delipidation than energisation but was, nevertheless, strongly inhibited at 50 percent lipid depletion. Spin label studies indicated a decrease in the fluidity of particle membranes accompanying delipidation. Moreover, the discontinuity seen in Arrhenius plots of ATPase activity was shifted from 17 degrees C (control) to 22 degrees C at 50 percent phospholipid depletion. The data are consistent with a release of unsaturated fatty acids by phospholipase A2 rendering the transport particles both leakier and the membranes less fluid than controls.
D000818:Animals; D002417:Cattle; D004578:Electron Spin Resonance Spectroscopy; D004579:Electron Transport / Q000187:drug effects*; D007425:Intracellular Membranes / Q000187:drug effects / Q000378:metabolism; D008563:Membrane Lipids / Q000378:metabolism*; D008564:Membrane Potentials / Q000187:drug effects; D008565:Membrane Proteins / Q000378:metabolism; D008929:Mitochondria, Heart / Q000187:drug effects / Q000378:metabolism; D010740:Phospholipases / Q000494:pharmacology*; D010741:Phospholipases A / Q000378:metabolism / Q000494:pharmacology*; D054467:Phospholipases A2; D010743:Phospholipids / Q000378:metabolism*; D006180:Proton-Translocating ATPases / Q000378:metabolism*; D011522:Protons
D008563:Membrane Lipids; D008565:Membrane Proteins; D010743:Phospholipids; D011522:Protons; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2; D006180:Proton-Translocating ATPases