ATP synthase (EC 3.6.3.14) is an important enzyme that provides energy for the cell to use through the synthesis of adenosine triphosphate (ATP). ATP is the most commonly used "energy currency" of cells from most organisms. It is formed from adenosine diphosphate (ADP) and inorganic phosphate (Pi) which releases energy.
The overall reaction sequence is: ATP synthase + ADP + Pi → ATP Synthase + ATP
This energy is often in the form of protium or H+, moving down an electrochemical gradient, such as from the lumen into the stroma of chloroplasts or from the inter-membrane space into the matrix in mitochondria.
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Located within the mitochondria ATP synthase consists of 2 regions
The nomenclature of the enzyme suffers from a long history. The F1 fraction derives its name from the term "Fraction 1" and FO (written as a subscript letter "o", not "zero") derives its name from being the oligomycin binding fraction.[1] Oligomycin, an antibiotic, is able to inhibit the FO unit of ATP synthase.
These functional regions consist of different protein subunits - refer to tables.
The F1 particle is large and can be seen in the transmission electron microscope by negative staining.[2] 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.
| Subunit | Human Gene |
|---|---|
| alpha | ATP5A1, ATPAF2 |
| beta | ATP5B, ATPAF1, C16orf7 |
| gamma | ATP5C1 |
| delta | ATP5D |
| epsilon | ATP5E |
The FO region of ATP synthase is a proton pore which is emmbeded into the mitochondrial membrane. It consists of three main subunits A, B and C, and (in humans) six additional subunits, d, e, f, g, F6 and 8 (or A6L).
| Subunit | Human Gene |
|---|---|
| A | ATP6 |
| B | ATP5F1 |
| C | ATP5G1, ATP5G2, ATP5G3 |
In the 1960s through the 1970s, Paul Boyer developed the 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 John E. Walker, then at the MRC Laboratory of Molecular Biology in Cambridge but now at the MRC Mitochondrial Biology Unit (also in Cambridge), crystallized the F1 catalytic-domain of ATP synthase. The structure, at the time the largest asymmetric protein structure known, indicated that Boyer's rotary-catalysis model was, in essence, correct. For elucidating this, Boyer and Walker shared half of the 1997 Nobel Prize in Chemistry. 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 subunits (3 of each), arranged like segments of an orange around an asymmetrical gamma subunit. According to the current model of ATP synthesis (known as the alternating catalytic model), the proton-motive force across the inner mitochondrial membrane, generated by the electron transport chain, drives the passage of protons through the membrane via the FO region of ATP synthase. A portion of the FO (the ring of c-subunits) rotates as the protons pass through the membrane. The c-ring is tightly attached to the asymmetric central stalk (consisting primarily of the gamma subunit), which rotates within the alpha3beta3 of F1 causing the 3 catalytic nucleotide binding sites to go through a series of conformational changes that leads to ATP synthesis. The major F1 subunits are prevented from rotating in sympathy with the central stalk rotor by a peripheral stalk that joins the alpha3beta3 to the non-rotating portion of FO. The structure of the intact ATP synthase is currently known at low-resolution from electron cryo-microscopy (cryo-EM) studies of the complex. The cryo-EM model of ATP synthase suggests that the peripheral stalk is a flexible structure that wraps around the complex as it joins F1 to FO. 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.[3] In the "open" state, ADP and phosphate enter the active site, in the diagram to the right this is shown in red. The protein then closes up around the molecules and binds them loosely - the "loose" state (shown in orange). 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.[4]
Like other enzymes, the activity of F1FO ATP synthase is reversible. Large-enough quantities of ATP cause it to create a transmembrane proton gradient, this is used by fermenting bacteria that 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, in general, runs in the opposite direction, creating ATP while using the protonmotive force created by the electron transport chain as a source of energy. The overall process of creating energy in this fashion is termed oxidative phosphorylation. The same process takes place in the mitochondria, where ATP synthase is located in the inner mitochondrial membrane (so that F1-part sticks into mitochondrial matrix, where ATP synthesis takes place).
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.[5][6] This coupling must have occurred early in the evolution of life as evidenced by essentially the same structure and processes of ATP synthase enzymes conserved in all kingdoms of life.[5] The F-ATP synthase shows large amounts of similarity both functionally and mechanically to the V-ATPase.[7] However whilst the F-ATP synthase generates ATP by utilising a proton gradient the V-ATPase is responsible for generating a proton gradient at the expense of ATP, generating pH values as low as 1. The F1 particle also shows significant similarity to hexameric DNA helicases and the FO particle shows some similarity to H+ powered flagellar motor complexes.[7] 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.[8]
The H+ motor of the FO particle shows great functional similarity to the H+ motors seen in flagellar motors.[7] Both feature a ring of many small alpha helical proteins that 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 FO particle and the ring of rotating proteins is far larger, with around 30 compared to the 10, 11, or 14 known in the FO 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.[5][8] This would then evolve to become more efficient, and eventually develop into the complex ATP synthases seen today. In alternative fashion, the DNA helicase/H+ motor complex may have had H+ pump activity, the ATPase activity of the helicase driving the H+ motor in reverse.[5] This could later evolve to carry out the reverse reaction and act as an ATP synthase.[6]
The following is a list of humans genes that encode components of ATP synthases:
In plants, ATP synthase is also present in chloroplasts (CF1FO-ATP synthase). The enzyme is integrated into thylakoid membrane; the CF1-part sticks into stroma, where dark reactions of photosynthesis (Also called the light-independent reactions or the Calvin cycle) and ATP synthesis take place. The overall structure and the catalytic mechanism of the chloroplast ATP synthase are almost the same as those of the mitochondrial enzyme. However, in chloroplasts, the proton motive force is generated not by respiratory electron transport chain but by primary photosynthetic proteins.
The ATP synthase isolated from bovine heart mitochondria (Bos taurus) is, in terms of biochemistry and structure, the best-characterized ATP synthase. Beef heart is used as a source for the enyzme because of the high concentration of mitochondria in cardiac muscle.
E. coli ATP synthase is the simplest known form of ATP synthase, with 8 different subunit types.
Yeast ATP synthase is one of the best-studied eukaryotic ATP synthases; and five F1, eight FO subunits, and seven associated proteins have been identified.[9] Most of these proteins have homologues in other eukaryotes.[10]
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