Redesigned and reversed: architectural and functional oddities of the trypanosomal ATP synthase

Gahura, Ondřej; Hierro-Yap, Carolina; Zı́ková, Alena

doi:10.1017/s0031182021000202

Cited by 21 publications

(35 citation statements)

References 110 publications

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“…The T. brucei ATP-synthase complex is composed of two subcomplexes, the membrane-embedded F o and the soluble F 1 , which are connected by a rotary central stalk and a stationary peripheral stalk [ 27 ]. The effect of drug treatment on the stability of the T. brucei ATP-synthase complex was investigated by exposing bloodstream forms to 50–400 nM compound 1 or 2 for 1 h, and extracts were separated by native PAGE ( Figure 9 A).…”

Section: Resultsmentioning

confidence: 99%

Cellular and Molecular Targets of Nucleotide-Tagged Trithiolato-Bridged Arene Ruthenium Complexes in the Protozoan Parasites Toxoplasma gondii and Trypanosoma brucei

Anghel

Müller

Serricchio

et al. 2021

IJMS

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Toxoplasma gondii is an apicomplexan parasite that infects and proliferates within many different types of host cells and infects virtually all warm-blooded animals and humans. Trypanosoma brucei is an extracellular kinetoplastid that causes human African trypanosomiasis and Nagana disease in cattle, primarily in rural sub-Saharan Africa. Current treatments against both parasites have limitations, e.g., suboptimal efficacy and adverse side effects. Here, we investigate the potential cellular and molecular targets of a trithiolato-bridged arene ruthenium complex conjugated to 9-(2-hydroxyethyl)-adenine (1), which inhibits both parasites with IC50s below 10−7 M. Proteins that bind to 1 were identified using differential affinity chromatography (DAC) followed by shotgun-mass spectrometry. A trithiolato-bridged ruthenium complex decorated with hypoxanthine (2) and 2-hydroxyethyl-adenine (3) were included as controls. Transmission electron microscopy (TEM) revealed distinct ultrastructural modifications in the mitochondrion induced by (1) but not by (2) and (3) in both species. DAC revealed 128 proteins in T. gondii and 46 proteins in T. brucei specifically binding to 1 but not 2 or 3. In T. gondii, the most abundant was a protein with unknown function annotated as YOU2. This protein is a homolog to the human mitochondrial inner membrane translocase subunit Tim10. In T. brucei, the most abundant proteins binding specifically to 1 were mitochondrial ATP-synthase subunits. Exposure of T. brucei bloodstream forms to 1 resulted in rapid breakdown of the ATP-synthase complex. Moreover, both datasets contained proteins involved in key steps of metabolism and nucleic acid binding proteins.

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Section: Resultsmentioning

confidence: 99%

Cellular and Molecular Targets of Nucleotide-Tagged Trithiolato-Bridged Arene Ruthenium Complexes in the Protozoan Parasites Toxoplasma gondii and Trypanosoma brucei

Anghel

Müller

Serricchio

et al. 2021

IJMS

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“…The dimers are sensitive to the lack of cardiolipin 14 and form short left-handed helical segments that extend across the membrane ridge of the discoidal cristae 15 . Uniquely among aerobic eukaryotes, the mammalian life cycle stage of T. brucei utilizes the ATP synthase as a proton pump maintaining the mitochondrial membrane potential at the expense of ATP 16,17 , whereas the insect stage of the parasite employs the forward ATP-producing mode of the enzyme 18,19 .…”

Section: Mainmentioning

confidence: 99%

An ancestral interaction module promotes oligomerization in divergent mitochondrial ATP synthases

Gahura

Muhleip

Hierro-Yap

et al. 2021

Preprint

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Mitochondrial ATP synthase forms stable dimers arranged into oligomeric assemblies that generate the inner-membrane curvature essential for efficient energy conversion. Here, we report cryo EM structures of the intact ATP synthase dimer from trypanosomes in 10 different rotational states. The model consists of 25 subunits, including 11 lineage-specific, as well as 36 lipids. The rotary mechanism is influenced by the divergent peripheral stalk, conferring a greater conformational flexibility. Proton transfer in the lumenal half-channel occurs via a chain of five ordered water molecules. The dimerization interface is formed by subunit-g that is critical for interactions but not for the catalytic activity. Although overall dimer architecture varies among eukaryotes, we find that subunit-g and -e form a common ancestral oligomerisation motif, which is shared between the trypanosomal and mammalian lineages. Therefore, our data defines the subunit-g/e module as a structural component determining ATP synthase oligomeric assemblies.

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“…Despite overall structural and functional conservation between F1Fo ATP synthases of all three domains (12), there is now evidence for substantial divergence between eukaryotic lineages in proteins making up the non-core parts of the complex (13)(14)(15)(16)(17)(18)(19)(20). Subunits of the peripheral stalk and subunits involved in the anchorage of the peripheral stalk to the membrane, in holding the Fo a subunit in proximity to the c-ring and in the dimerisation domains all show in general low sequence conservation between species.…”

Section: Introductionmentioning

confidence: 99%

“…Subunits of the peripheral stalk and subunits involved in the anchorage of the peripheral stalk to the membrane, in holding the Fo a subunit in proximity to the c-ring and in the dimerisation domains all show in general low sequence conservation between species. However, while many non-core ATPase subunits show no sequence similarity between species, structural homology can sometimes still be detected (13,14,21).…”

Section: Introductionmentioning

confidence: 99%

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Characterisation of a highly diverged mitochondrial ATP synthase peripheral stalk subunit b in Trypanosoma brucei

Dewar

Oeljeklaus²,

Warscheid

et al. 2021

Preprint

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The mitochondrial F1Fo ATP synthase of Trypanosoma brucei has been studied in detail. Whereas its F1 moiety is relatively highly conserved in structure and composition, the same is not the case for the Fo part and the peripheral stalk. A core subunit of the latter, the normally conserved subunit b, could not be identified in trypanosomes suggesting that it might be absent. Here we have identified a 17 kDa mitochondrial protein of the inner membrane that is essential for normal growth, efficient oxidative phosphorylation and membrane potential maintenance. Pulldown experiments and native PAGE analysis indicate that the protein is associated with the F1Fo ATP synthase. Its ablation reduces the levels of Fo subunits, but not those of F1, and disturbs the cell cycle. HHpred analysis showed that the protein has structural similarities to subunit b of other species, indicating that the Fo part of the trypanosomal ATP synthase does contain a highly diverged subunit b. Thus, the Fo part of the trypanosomal ATPase synthase may be more widely conserved than initially thought.

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Redesigned and reversed: architectural and functional oddities of the trypanosomal ATP synthase

Abstract: Abstract

Cited by 21 publications

References 110 publications

Cellular and Molecular Targets of Nucleotide-Tagged Trithiolato-Bridged Arene Ruthenium Complexes in the Protozoan Parasites Toxoplasma gondii and Trypanosoma brucei

Cellular and Molecular Targets of Nucleotide-Tagged Trithiolato-Bridged Arene Ruthenium Complexes in the Protozoan Parasites Toxoplasma gondii and Trypanosoma brucei

An ancestral interaction module promotes oligomerization in divergent mitochondrial ATP synthases

Characterisation of a highly diverged mitochondrial ATP synthase peripheral stalk subunit b in Trypanosoma brucei

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