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LIFE 210 (38)

Chemiosmotic theory, mitochondrial inner membrane vesicles, ATP synthasem cancer cell metabolism

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Colorado State University
Life Science
LIFE 210
Paul Laybourn

5 October Electrochemical Gradient Produced by ETC ETC Uses –ΔG of e transfer from NADH to O 2 Transfers H out of matrix Electrochemical gradient across IM Membrane 2 components Charge ΔV or ΔΨ = 140 mV ~70% of –ΔG pH + ΔpH or Δ[H ] = 60 mV ~30% of –ΔG -ΔG = ΔP or pmf = -ΔpH + ΔΨ = -1.4 kcal/mol + -3.2 kcal/mol Total –ΔG = -4.6 kcal/mol Peter Mitchell & the Chemiosmotic Theory In 1961 Radical idea at the time + Energy from NADH & FADH stor2d as an electrochemical H gradient Used to drive ATP synthesis Before then it was assumed that all ATP synthesis was through substrate-level phosphorylation Mitchell received the Nobel Prize in Chemistry in 1978 Mitochondrial Inner Membrane Vesicles (MIMs) and AtP Synthase Mitochondrian + mild detergent takes off outer membrane Left with inner mitochondrial membrane Sonicate → breaks off bits of membranes and forms vesicles Some vesicles are inside out – makes important parts accessible (e.g. ATP synthase) Have ETC & ATP synthase + Use to show H gradient is required Add dinitrophenol (uncoupler) + Membrane soluble H carrier Uncouples ETC from ATP synthase Uncoupler shuttles protons out – no gradient Just produces heat Brown adipose fall cells Found in infants/hibernating mammals Functions to produce heat Nonshivering thermogenesis Too cold to shiver, still produces heat Use to show H gradient is sufficient Soak MIMs at pH 4 Leads to high [H ] inside Transfer to pH 8 Creates concentration gradient Add ADP and Pi → ATP created + NADH & O not2required if there is another way to form an H gradient
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