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oai:figshare.com:article/34025422 2026-09-29T17:22:10Z category_4 category_8 category_12 category_13 category_24 category_272 category_734 portal_5 item_type_1 month_year_09_2026

Analysis of ATP synthase. Andrew E. Maclean (10219517) Orsola Iorillo (22772687) Drahomíra Faktorová (373414) Monika Singh (593097) Suzanne McGill (11916067) Julius Lukeš (122896) Lilach Sheiner (79023) Biochemistry Microbiology Cell Biology Genetics Evolutionary Biology Environmental Sciences not elsewhere classified Biological Sciences not elsewhere classified shares new components mitoribosome possesses many div >< p highly conserved feature perkinsus marinus </ divergent subunit composition divergent mitochondrial complexes perkinsus </ conserved across toxoplasma </ plasmodium </ mitochondrial complexes metc complexes mitochondrial ribosome mitochondrial function xlink "> wider lineages unexpected diversity transcription factors study reveals outer membrane observed enlarged myzozoan parasite metc ), including members deadly apicomplexan complexome profiling cell survival atp synthase apiap2 family (A) Heatmap of the whole-cell complexome of ATP synthase. Each row represents a gel slice, with the gel slice number indicated on the left and the molecular weight, based on the migration of soluble proteins (blue), on the right. Subunit IDs are shown at the bottom of their corresponding profile. Dark blue indicates the highest relative abundance (1) and white the lowest (0). Coomassie-stained Native PAGE gel on which the analysis was performed is depicted on the left of the profile. (B) Abundance profiles of ATPPM1, ATPPM2, and the average abundance of all ATPTG subunits across all gel slices. Protein abundance is displayed as iBAQ values. Data shown is from replicate 1. (C) Chromatogram displaying UV 1_280 of gel filtration of purified ATP synthase using a Superose 6 increase 3.2/300 column. Left-most peak represents void-volume. The fractions highlighted in blue were combined and concentrated using a 100 kDa MWCO spin concentrator before analysis via immunoblot and mass spectrometry. (D) SDS-PAGE analysis of purified ATP synthase. Gel was either silver stained to visualise protein subunits or subjected to immunoblot analysis with the ATPβ antibody. (E) Structural prediction of P. marinus ATPTG1 (predicted using Alphafold3 [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1014543#ppat.1014543.ref100" target="_blank">100</a>]), pTM score 0.41, coloured according to pIDDT score (left), the structure of T. gondii ATPTG1 (PDB 6TMK; [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1014543#ppat.1014543.ref017" target="_blank">17</a>]) (center) and the superimposition of the two structures (RMSD = 1.246 Å) (right). Visualised using ChimeraX [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1014543#ppat.1014543.ref098" target="_blank">98</a>]. (F) Alignment of ATPTG1, CytC1 and fusion proteins from across the myzozoa. ATPTG1 proteins: Perkinsus marinus Pmar_PMAR013021; Chromera velia Cvel_13300; Vitrella brassicaformis Vbra_20035; Cryptosporidium muris CMU_009920. CytC1 proteins: Perkinsus marinus Pmar_PMAR028703; Vitrella brassicaformis Vbra_21749. Fusion proteins: TGGT1_246540 from Toxoplasma gondii; PF3D7_1462700 from Plasmodium falciparum. Alignments were made using Clustal Omega and visualised using JalView [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1014543#ppat.1014543.ref099" target="_blank">99</a>]. Colour coding depicts percent identity. (TIF) 2026-09-29T17:22:05Z Image Figure 10.1371/journal.ppat.1014543.s003 https://figshare.com/articles/figure/_p_Analysis_of_ATP_synthase_p_/34025422 CC BY 4.0