Plastid terminal oxidase

Last updated
Plastid terminal oxidase
Identifiers
SymbolPTOX
Pfam PF01786
Available protein structures:
Pfam   structures / ECOD  
PDB RCSB PDB; PDBe; PDBj
PDBsum structure summary

Plastid terminal oxidase or plastoquinol terminal oxidase (PTOX) is an enzyme that resides on the thylakoid membranes of plant and algae chloroplasts and on the membranes of cyanobacteria. The enzyme was hypothesized to exist as a photosynthetic oxidase in 1982 and was verified by sequence similarity to the mitochondrial alternative oxidase (AOX). [1] The two oxidases evolved from a common ancestral protein in prokaryotes, and they are so functionally and structurally similar that a thylakoid-localized AOX can restore the function of a PTOX knockout. [2]

Contents

Function

Plastid terminal oxidase catalyzes the oxidation of the plastoquinone pool, which exerts a variety of effects on the development and functioning of plant chloroplasts.

A summary of the pathways plastid terminal oxidase plays a role in through oxidation of the quinone pool Role of plastid terminal oxidase.png
A summary of the pathways plastid terminal oxidase plays a role in through oxidation of the quinone pool

Carotenoid biosynthesis and plastid development

The enzyme is important for carotenoid biosynthesis during chloroplast biogenesis. In developing plastids, its activity prevents the over-reduction of the plastoquinone pool. Knockout plants for PTOX exhibit phenotypes of variegated leaves with white patches. Without the enzyme, the carotenoid synthesis pathway slows down due to the lack of oxidized plastoquinone with which to oxidize phytoene, a carotenoid intermediate. The colorless compound phytoene accumulates in the leaves, resulting in white patches of cells. [3] PTOX is also thought to determine the redox poise of the developing photosynthetic apparatus and without it, plants fail to assemble organized internal membrane structures in chloroplasts when exposed to high light during early development. [1] [4]

Photoprotection

Plants deficient in the IMMUTANS gene that encodes the oxidase are especially susceptible to photooxidative stress during early plastid development. The knockout plants exhibit a phenotype of variegated leaves with white patches that indicate a lack of pigmentation or photodamage. This effect is enhanced with increased light and temperature during plant development. The lack of plastid terminal oxidase indirectly causes photodamage during plastid development because protective carotenoids are not synthesized without the oxidase. [5]

The enzyme is also thought to act as a safety valve for stress conditions in the photosynthetic apparatus. By providing an electron sink when the plastoquinone pool is over-reduced, the oxidase is thought to protect photosystem II from oxidative damage. Knockouts for Rubisco and photosystem II complexes, which would experience more photodamage than normal, exhibit an upregulation of plastid terminal oxidase. [6] This effect is not universal because it requires plants to have additional PTOX regulation mechanisms. While many studies agree with the stress-protective role of the enzyme, one study showed that overexpression of PTOX increases the production of reactive oxygen species and causes more photodamage than normal. This finding suggests that an efficient antioxidant system is required for the oxidase to function as a safety valve for stress conditions and that it is more important during chloroplast biogenesis than in the regular functioning of the chloroplast. [7]

Chlororespiration and electron flux

The most confirmed function of plastid terminal oxidase in developed chloroplasts is its role in chlororespiration. In this process, NADPH dehydrogenase (NDH) reduces the quinone pool and the terminal oxidase oxidizes it, serving the same function as cytochrome c oxidase from mitochondrial electron transport. In Chlamydomonas , there are two copies of the gene for the oxidase. PTOX2 significantly contributes to the flux of electrons through chlororespiration in the dark. [8] There is also evidence from experiments with tobacco that it functions in plant chlororespiration as well. [9]

In fully developed chloroplasts, prolonged exposure to light increases the activity of the oxidase. Because the enzyme acts at the plastoquinone pool in between photosystem II and photosystem I, it may play a role in controlling electron flow through photosynthesis by acting as an alternative electron sink. Similar to its role in carotenoid synthesis, its oxidase activity may prevent the over-reduction of photosystem I electron acceptors and damage by photoinhibition. A recent analysis of electron flux through the photosynthetic pathway shows that even when activated, the electron flux plastid terminal oxidase diverts is two orders of magnitude less than the total flux through photosynthetic electron transport. [10] This suggests that the protein may play less of a role than previously thought in relieving the oxidative stress in photosynthesis.

Structure

Plastid terminal oxidase is an integral membrane protein, or more specifically, an integral monotopic protein and is bound to the thylakoid membrane facing the stroma. Based on sequence homology, the enzyme is predicted to contain four alpha helix domains that encapsulate a di-iron center. The two iron atoms are ligated by six essential conserved histidine and glutamate residues – Glu136, Glu175, His171, Glu227, Glu296, and His299. [11] The predicted structure is similar to that of the alternative oxidase, with an additional Exon 8 domain that is required for the plastid oxidase's activity and stability. The enzyme is anchored to the membrane by a short fifth alpha helix that contains a Tyr212 residue hypothesized to be involved in substrate binding. [12]

Mechanism

The oxidase catalyzes the transfer of four electrons from reduced plastoquinone to molecular oxygen to form water . The net reaction is written below:

2 QH2 + O2 → 2 Q + 2 H2O

Analysis of substrate specificity revealed that the enzyme almost exclusively catalyzes the reduction of plastoquinone over other quinones such as ubiquinone and duroquinone. Additionally, iron is essential for the catalytic function of the enzyme and cannot be substituted by another metal cation like Cu2+, Zn2+, or Mn2+ at the catalytic center. [13]

It is unlikely that four electrons could be transferred at once in a single iron cluster, so all of the proposed mechanisms involve two separate two-electron transfers from reduced plastoquinone to the di-iron center. In the first step common to all proposed mechanisms, one plastoquinone is oxidized and both irons are reduced from iron(III) to iron(II). Four different mechanisms are proposed for the next step, oxygen capture. One mechanism proposes a peroxide intermediate, after which one oxygen atom is used to create water and another is left bound in a diferryl configuration. Upon one more plastoquinone oxidation, a second water molecule is formed and the irons return to a +3 oxidation state. The other mechanisms involve the formation of Fe(III)-OH or Fe(IV)-OH and a tyrosine radical. [14] These radical-based mechanisms could explain why over-expression of the PTOX gene causes increased generation of reactive oxygen species.

Evolution

The enzyme is present in organisms capable of oxygenic photosynthesis, which includes plants, algae, and cyanobacteria. Plastid terminal oxidase and alternative oxidase are thought to have originated from a common ancestral di-iron carboxylate protein. Oxygen reductase activity was likely an ancient mechanism to scavenge oxygen in the early transition from an anaerobic to aerobic world. The plastid oxidase first evolved in ancient cyanobacteria and the alternative oxidase in Pseudomonadota before eukaryotic evolution and endosymbiosis events. Through endosymbiosis, the plastid oxidase was vertically inherited by eukaryotes that evolved into plants and algae. Sequenced genomes of various plant and algae species shows that the amino acid sequence is more than 25% conserved, which is a significant amount of conservation for an oxidase. This sequence conservation further supports the theory that both the alternative and plastid oxidases evolved before endosymbiosis and did not significantly change through eukaryote evolution. [15]

There also exist PTOX cyanophages that contain copies of the gene for the plastid oxidase. They are known to act as viral vectors for movement of the gene between cyanobacterial species. Some evidence suggests that the phages may use the oxidase to influence photosynthetic electron flow to produce more ATP and less NADPH because viral synthesis utilizes more ATP. [1]

See also

Related Research Articles

<span class="mw-page-title-main">Chloroplast</span> Plant organelle that conducts photosynthesis

A chloroplast is a type of membrane-bound organelle known as a plastid that conducts photosynthesis mostly in plant and algal cells. The photosynthetic pigment chlorophyll captures the energy from sunlight, converts it, and stores it in the energy-storage molecules ATP and NADPH while freeing oxygen from water in the cells. The ATP and NADPH is then used to make organic molecules from carbon dioxide in a process known as the Calvin cycle. Chloroplasts carry out a number of other functions, including fatty acid synthesis, amino acid synthesis, and the immune response in plants. The number of chloroplasts per cell varies from one, in unicellular algae, up to 100 in plants like Arabidopsis and wheat.

<span class="mw-page-title-main">Oxidative phosphorylation</span> Metabolic pathway

Oxidative phosphorylation or electron transport-linked phosphorylation or terminal oxidation is the metabolic pathway in which cells use enzymes to oxidize nutrients, thereby releasing chemical energy in order to produce adenosine triphosphate (ATP). In eukaryotes, this takes place inside mitochondria. Almost all aerobic organisms carry out oxidative phosphorylation. This pathway is so pervasive because it releases more energy than alternative fermentation processes such as anaerobic glycolysis.

<span class="mw-page-title-main">Photosynthesis</span> Biological process to convert light into chemical energy

Photosynthesis is a biological process used by many cellular organisms to convert light energy into chemical energy, which is stored in organic compounds that can later be metabolized through cellular respiration to fuel the organism's activities. The term usually refers to oxygenic photosynthesis, where oxygen is produced as a byproduct and some of the chemical energy produced is stored in carbohydrate molecules such as sugars, starch, glycogen and cellulose, which are synthesized from endergonic reaction of carbon dioxide with water. Most plants, algae and cyanobacteria perform photosynthesis; such organisms are called photoautotrophs. Photosynthesis is largely responsible for producing and maintaining the oxygen content of the Earth's atmosphere, and supplies most of the biological energy necessary for complex life on Earth.

<span class="mw-page-title-main">Symbiogenesis</span> Evolutionary theory holding that eukaryotic organelles evolved through symbiosis with prokaryotes

Symbiogenesis is the leading evolutionary theory of the origin of eukaryotic cells from prokaryotic organisms. The theory holds that mitochondria, plastids such as chloroplasts, and possibly other organelles of eukaryotic cells are descended from formerly free-living prokaryotes taken one inside the other in endosymbiosis. Mitochondria appear to be phylogenetically related to Rickettsiales bacteria, while chloroplasts are thought to be related to cyanobacteria.

<span class="mw-page-title-main">Thylakoid</span> Membrane enclosed compartments in chloroplasts and cyanobacteria

Thylakoids are membrane-bound compartments inside chloroplasts and cyanobacteria. They are the site of the light-dependent reactions of photosynthesis. Thylakoids consist of a thylakoid membrane surrounding a thylakoid lumen. Chloroplast thylakoids frequently form stacks of disks referred to as grana. Grana are connected by intergranal or stromal thylakoids, which join granum stacks together as a single functional compartment.

<span class="mw-page-title-main">Chromoplast</span> Pigment-bearing organelle in plant cells

Chromoplasts are plastids, heterogeneous organelles responsible for pigment synthesis and storage in specific photosynthetic eukaryotes. It is thought that like all other plastids including chloroplasts and leucoplasts they are descended from symbiotic prokaryotes.

<span class="mw-page-title-main">Plastoquinone</span> Molecule which moves electron in photosynthesis

Plastoquinone (PQ) is an isoprenoid quinone molecule involved in the electron transport chain in the light-dependent reactions of photosynthesis. The most common form of plastoquinone, known as PQ-A or PQ-9, is a 2,3-dimethyl-1,4-benzoquinone molecule with a side chain of nine isoprenyl units. There are other forms of plastoquinone, such as ones with shorter side chains like PQ-3 as well as analogs such as PQ-B, PQ-C, and PQ-D, which differ in their side chains. The benzoquinone and isoprenyl units are both nonpolar, anchoring the molecule within the inner section of a lipid bilayer, where the hydrophobic tails are usually found.

<span class="mw-page-title-main">Photosystem</span> Structural units of protein involved in photosynthesis

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<span class="mw-page-title-main">Photosystem II</span> First protein complex in light-dependent reactions of oxygenic photosynthesis

Photosystem II is the first protein complex in the light-dependent reactions of oxygenic photosynthesis. It is located in the thylakoid membrane of plants, algae, and cyanobacteria. Within the photosystem, enzymes capture photons of light to energize electrons that are then transferred through a variety of coenzymes and cofactors to reduce plastoquinone to plastoquinol. The energized electrons are replaced by oxidizing water to form hydrogen ions and molecular oxygen.

<span class="mw-page-title-main">Photosystem I</span> Second protein complex in photosynthetic light reactions

Photosystem I is one of two photosystems in the photosynthetic light reactions of algae, plants, and cyanobacteria. Photosystem I is an integral membrane protein complex that uses light energy to catalyze the transfer of electrons across the thylakoid membrane from plastocyanin to ferredoxin. Ultimately, the electrons that are transferred by Photosystem I are used to produce the moderate-energy hydrogen carrier NADPH. The photon energy absorbed by Photosystem I also produces a proton-motive force that is used to generate ATP. PSI is composed of more than 110 cofactors, significantly more than Photosystem II.

Cytochrome b<sub>6</sub>f complex Enzyme

The cytochrome b6f complex (plastoquinol/plastocyanin reductase or plastoquinol/plastocyanin oxidoreductase; EC 7.1.1.6) is an enzyme found in the thylakoid membrane in chloroplasts of plants, cyanobacteria, and green algae, that catalyzes the transfer of electrons from plastoquinol to plastocyanin:

<span class="mw-page-title-main">Photosynthetic reaction centre</span> Molecular unit responsible for absorbing light in photosynthesis

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<span class="mw-page-title-main">Photoinhibition</span>

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<span class="mw-page-title-main">Alternative oxidase</span>

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<span class="mw-page-title-main">Photosynthetic reaction centre protein family</span>

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<span class="mw-page-title-main">Light-dependent reactions</span> Photosynthetic reactions

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<span class="mw-page-title-main">15-Cis-phytoene desaturase</span> Class of enzymes

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<span class="mw-page-title-main">Chlororespiration</span>

Chlororespiration is a respiratory process that takes place within plants. Inside plant cells there is an organelle called the chloroplast which is surrounded by the thylakoid membrane. This membrane contains an enzyme called NAD(P)H dehydrogenase which transfers electrons in a linear chain to oxygen molecules. This electron transport chain (ETC) within the chloroplast also interacts with those in the mitochondria where respiration takes place. Photosynthesis is also a process that Chlororespiration interacts with. If photosynthesis is inhibited by environmental stressors like water deficit, increased heat, and/or increased/decreased light exposure, or even chilling stress then chlororespiration is one of the crucial ways that plants use to compensate for chemical energy synthesis.

References

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