ATP synthase

Structure and nomenclature

In mitochondria, the F₁F_O ATP synthase has a long history of scientific study.

• the F_O portion is within the membrane.
• The F₁ portion of the ATP synthase is above the membrane.

It's easy to visualize the F_OF₁ particle as resembling the fruiting body of a common mushroom, with the head being the F₁ particle, the stalk being the gamma subunit of F₁, and the base and "roots" being the F_O particle embedded in the membrane.

The nomenclature of the enzyme suffers from a long history. The F₁ fraction derives it name from the term "Fraction 1" and F_O (written as a subscript "O", not "zero") derives its name from being the oligomycin binding fraction.

Taking as an example the nomenclature of subunits in the bovine enzyme, many subunits have alphabet names:

• Greek letters: alpha, beta, gamma, delta, epsilon
• Roman letters: a, b, c, d, e, f, g, h

Others have more complex names:

• F₆ (from "Fraction 6")
• OSCP (the oligomycin sensitivity conferral protein), ATP5O
• A6L (named for the gene that codes for it in the mitochondrial genome)
• IF1 (inhibitory factor 1), ATPIF1

The F₁ particle is large and can be seen in the transmission electron microscope by negative staining.^[1] These are particles of 9 nm diameter that pepper the inner mitochondrial membrane. They were originally called elementary particles and were thought to contain the entire respiratory apparatus of the mitochondrion, but through a long series of experiments, Ephraim Racker and his colleagues (who first isolated the F₁ particle in 1961) were able to show that this particle is correlated with ATPase activity in uncoupled mitochondria and with the ATPase activity in submitochondrial particles created by exposing mitochondria to ultrasound. This ATPase activity was further associated with the creation of ATP by a long series of experiments in many laboratories.

Binding change mechanism

In the 1960s through the 1970s, Paul Boyer developed his binding change, or flip-flop, mechanism, which postulated that ATP synthesis is coupled with a conformational change in the ATP synthase generated by rotation of the gamma subunit. The research group of Jens Christian Skou received the other half of the Chemistry prize that year "for the first discovery of an ion-transporting enzyme, Na+, K+ -ATPase"  

The crystal structure of the F₁ showed alternating alpha and beta electron cryo-microscopy (cryo-EM) studies of the complex. The cryo-EM model of ATP synthase shows that the peripheral stalk is a flexible rope-like structure that wraps around the complex as it joins F₁ to F_O. Under the right conditions, the enzyme reaction can also be carried out in reverse, with ATP hydrolysis driving proton pumping across the membrane.

The binding change mechanism involves the active site of a β subunit cycling between three states.^[2] In the "open" state, ADP and phosphate enter the active site, in the diagram to the right this is shown in brown. The protein then closes up around the molecules and binds them loosely - the "loose" state (shown in red). The enzyme then undergoes another change in shape and forces these molecules together, with the active site in the resulting "tight" state (shown in pink) binding the newly-produced ATP molecule with very high affinity. Finally, the active site cycles back to the open state, releasing ATP and binding more ADP and phosphate, ready for the next cycle of ATP production.

Physiological role

Like other enzymes, the activity of F₁F_O ATP synthase is reversible. Large enough quantities of ATP cause it to create a transmembrane proton gradient, this is used by fermenting bacteria which do not have an electron transport chain, and hydrolyze ATP to make a proton gradient, which they use for flagella and transport of nutrients into the cell.

In respiring bacteria under physiological conditions, ATP synthase generally runs in the opposite direction, creating ATP while using the oxidative phosphorylation. The same process takes place in mitochondria, where ATP synthase is located in the inner mitochondrial membrane (so that F₁-part sticks into mitochondrial matrix, where ATP synthesis takes place).

ATP synthase in different organisms

Plant ATP synthase

In plants ATP synthase is also present in chloroplasts (CF₁F_O-ATP synthase). The enzyme is integrated into proton motive force is generated not by respiratory electron transport chain, but by primary photosynthetic proteins.

E. coli ATP synthase

E. coli ATP synthase is the simplest known form of ATP synthase, with 8 different subunit types.

Yeast ATP synthase

Yeast ATP synthase is the most complex known and is made of 20 different types of subunits.

Evolution of ATP synthase

The evolution of ATP synthase is thought to be an example of modular evolution, where two subunits with their own functions have become associated and gained new functionality. The F₁ particle shows significant similarity to hexameric DNA helicases and the F_O particle shows some similarity to H⁺ powered flagellar motor complexes.

The α₃β₃ hexamer of the F₁ particle shows significant structural similarity to hexameric DNA helicases; both form a ring with 3 fold rotational symmetry with a central pore. Both also have roles dependent on the relative rotation of a macromolecule within the pore; the DNA helicases use the helical shape of DNA to drive their motion along the DNA molecule and to detect supercoiling whilst the α₃β₃ hexamer uses the conformational changes due rotation of the γ subunit to drive an enzymatic reaction.

The H⁺ motor of the F_O particle shows great functional similarity to the H⁺ motors seen in flagellar motors. Both feature a ring of many small alpha helical proteins which rotate relative to nearby stationary proteins using a H⁺ potential gradient as an energy source. This is, however, a fairly tenuous link - the overall structure of flagellar motors is far more complex than the F_O particle and the ring of rotating proteins is far larger, with around 30 compared to the 10, 11 or 14 known in the F_O complex.

The modular evolution theory for the origin of ATP synthase suggests that two subunits with independent function, a DNA helicase with ATPase activity and a H⁺ motor, were able to bind, and the rotation of the motor drive the ATPase activity of the helicase in reverse. This would then evolve to become more efficient, and eventually develop into the complex ATP synthases seen today. Alternatively the DNA helicase/H⁺ motor complex may have had H⁺ pump activity, the ATPase activity of the helicase driving the H⁺ motor in reverse. This could later evolve to carry out the reverse reaction and act as an ATP synthase.

See also

• Oxidative phosphorylation
• Mitochondrion
• Chloroplast
• Electron transfer chain
• Proton pump
• Transmembrane ATPase
• Flavoprotein
• ATP synthase subunit C
• ATP synthase alpha/beta subunits
• P-ATPase

References

1. ^ Fernandez-Moran et al., Journal of Molecular Biology, Vol 22, p 63, 1962
2. ^ Gresser MJ, Myers JA, Boyer PD (1982). "Catalytic site cooperativity of beef heart mitochondrial F1 adenosine triphosphatase. Correlations of initial velocity, bound intermediate, and oxygen exchange measurements with an alternating three-site model". J. Biol. Chem. 257 (20): 12030-8. PMID 6214554.

v • d • e Acid anhydride hydrolases: Myosin

v • d • e Complex IV (3 units) - ATP synthase (2 units)