Fl particles have ATP synthase activity F spheres Inner mitochondrial (b)Submitochondrial particles membrane Electron transport? Yes ATP synthesis? Yes Outer mitochondrial ATPase activity/? No (a)Intact mitochondrion Electron transport? Yes ATP synthesis? Yes (c)Dissociated particles Electron transport? Yes ATP synthesis? No ○ ATPase activity? Yes (d)Membranous fraction 9 a(e)Soluble fraction with F, spheres Electron transport? Yes Electron transport? No ATP synthesis? No ATP synthesis? No ATPase activity? No ATPase activity? Yes (f)Reconstituted particles Electron transport? Yes ATP synthesis? Yes ATPase activity? No
vF1 particles have ATP synthase activity
.o Proton translocation through Fo drives atP synthesis by Fi: Binding change model and rotational catalysis ④4 Cycle begins again Loose subunit with knob rotated binds ADP+ P by one subunit Boyer proposed in ADP 1979 and was F knob, top view greatly stimulated ADP+ P by the publication T L in 1994 of the 60° rotation a structure for F1 complex(x-ray) H2o from bovine heart tochondria New ATP forms by (2 Energy input alters condensation; original ATP is released: L subunit available for binding
v Proton translocation through F0 drives ATP synthesis by F1: Binding Change Model and rotational catalysis Boyer proposed in 1979, and was greatly stimulated by the publication in 1994 of the structure for F1 complex (X-ray) from bovine heart mitochondria
o Direct experimental evidence supporting the rotational catalysis. Actin filament Japan researcher, a3B3y B ature 386: 300 complex/a 1997 His-tag Coverslip coated with Ni-NTA
vDirect experimental evidence supporting the rotational catalysis. Japan researcher, Nature 386: 300, 1997
o The atp synthase is a reversible coupling device H H H H H H H Inner H H mitochondrial H H membrane MATRIX H SPACE H H AD H ADP P ATP (A) ATP SYNTHESIS (B) ATP HYDROLYSIS
v The ATP synthase is a reversible coupling device
.o Other roles for the proton-motive force in addition to ATP synthase ADP3 ATP4 outer membrane voltage gradient drives ADP-ATP exchange inner membrane ADP3 ATP ATP4 ADP3 pyruvate H MATRIX pH gradient pyruvate pH gradient drives pyruvate drives phosphate Import import pyruvate ⊙
v Other roles for the proton-motive force in addition to ATP synthase