<p>Analysis of ATP synthase.</p>

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 <i>P. marinus</i> ATPTG1 (predicted using Alphafold3 [<a href="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 <i>T. gondii</i> ATPTG1 (PDB 6TMK; [<a href="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="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: <i>Perkinsus marinus</i> Pmar_PMAR013021; <i>Chromera velia</i> Cvel_13300<i>; Vitrella brassicaformis Vbra_20035</i>; <i>Cryptosporidium muris</i> CMU_009920.

CytC1 proteins: <i>Perkinsus marinus</i> Pmar_PMAR028703; <i>Vitrella brassicaformis</i> Vbra_21749. Fusion proteins: TGGT1_246540 from <i>Toxoplasma gondii</i>; PF3D7_1462700 from <i>Plasmodium falciparum</i>. Alignments were made using Clustal Omega and visualised using JalView [<a href="plospathogens.org/article/info:doi/10.1371/journal.ppat.1014543#ppat.1014543.ref099" target="_blank">99</a>].

Colour coding depicts percent identity.</p> <p>(TIF)</p>