Data · dataset · 2026
<p>Complexome profiles of mETC complexes.</p>
Listed in ZivaHub
<p>(A) Immunoblot analysis of total cell lysate (Total) and purification fractions using antibodies against marker proteins of various cell compartments.
Description
The pellet from the 3,500 x <i>g</i> centrifugation step contains unbroken cells and heavy cell components, from the 40,000 x <i>g</i> step the supernatant contains soluble cytosolic proteins, and the pellet mitochondrial and organellar material. 7.5 μg of protein was loaded per lane with equal loading confirmed by instant blue staining (bottom panel).
The following antibodies were used: ATPβ (mitochondrion), BiP (ER), GAPDH (cytosol) and Histone H3 (nucleus). (B) Blue-native PAGE analysis of mitochondrially-enriched material treated with βDDM and separated on a 4–16% Bis-Tris gel. Total protein (TP) was visualised by Coomassie Brilliant Blue staining (CBB), complex II (CII) activity by an in-gel succinate: nitroblue tetrazolium (NBT) activity stain, complex IV (CIV) activity by an in-gel cytochrome <i>c</i>: 3,3′-Diaminobenzidine (DAB) activity stain and ATP synthase (AS) by immunoblot analysis using the anti-ATPβ antibody.
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(C) Heatmaps of the complexome profiles from mitochondrially-enriched material of mETC complex subunits from complex II, complex III and complex IV. Each row represents a gel slice, with the gel slice number indicated on the left and the molecular weight, based on the migration of mammalian mitochondrial complexes (black) and soluble proteins (blue), on the right. Subunit IDs are shown at the bottom of their corresponding profile, with mitochondrially-encoded subunits highlighted in red.
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. Data shown is from replicate 1.
(D) Coulson plots depicting the composition of mETC complexes in key species from myzozoans and ciliates. <i>Perkinsus</i> subunits were used for homology searches using BLAST. <i>T.t Tetrahymena thermophila</i>; <i>P.t Paramecium tetraurelia</i>; <i>T.g Toxoplasma gondii</i>; <i>P.f Plasmodium falciparum</i>; <i>C.p Cryptosporidium parvum</i>; <i>C.m Cryptosporidium muris</i>; <i>C.v Chromera velia</i>; <i>V.b Vitrella brassicaformis</i>; <i>P.g Polarella glacialis</i>; <i>S.m Symbiodinium microadriaticum</i>.
Full dataset in <a href="plospathogens.org/article/info:doi/10.1371/journal.ppat.1014543#ppat.1014543.s011" target="_blank">S6 Table</a>. (E) Abundance profile of complex II subunit SDHG, complex III subunit RIP1 and complex IV subunit Cox2B across all gel slices. Protein abundance is displayed as iBAQ values.
Data shown is from whole-cell complexome. (F) Abundance profile of ISPR1, ISPR2 and complex III subunit RIP1 across all slices. Protein abundance is displayed as iBAQ values.
Data shown is from replicate 2. (G) Immunofluorescence assay of <i>Tg</i>ISPR. Samples labelled with anti-Ty to detect <i>Tg</i>ISPR-Ty (cyan) and anti-Tom40 (magenta) to detect the mitochondrial marker protein Tom40.
Scale bar is 2 µm. Plot to the right displays fluorescence profiles of Ty (cyan) and Tom40 (magenta) labelling along the white dotted line on the merged image.</p>
Links
Where it is published
- DOI doi.org/10.1371/journal.ppat.1014543.g002 ↗
DOI / persistent id · from zivahub uct ac za
Catalogue records · 1
- OAI-PMH record api.figshare.com/v2/oai?verb=GetRecord&metadataPrefix=oai_dc&identifier=oai%3Af… ↗
metadata API · from zivahub uct ac za
Topics
- From keywords
- Biochemistry and cell biology · Earth & Environmental Science · Evolutionary biology · Genetics · Life Sciences · Microbiology
Provenance · 1 source records, 13 field assertions
| Source | Key | Last seen | Raw |
|---|---|---|---|
| ZivaHub | oai:figshare.com:article/34025464 | 13 d ago | JSON v1 |
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| concepts[field].anzsrc:group:3105 | mapping · zivahub uct ac za | vocabulary-mapper@1.0.0 | keywords['Genetics'] |
| concepts[field].anzsrc:group:3107 | mapping · zivahub uct ac za | vocabulary-mapper@1.0.0 | keywords['Microbiology'] |
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| concepts[modality].local:modality:tabular | enrichment · zivahub uct ac za | keyword-concept-rules@1.0.0 | title+description (65%) |
| description | source · zivahub uct ac za | connector:zivahub_uct_ac_za@1.0.0 | /metadata/dc/description |
| license | source · zivahub uct ac za | connector:zivahub_uct_ac_za@1.0.0 | /metadata/dc/rights |
| publication_date | source · zivahub uct ac za | connector:zivahub_uct_ac_za@1.0.0 | |
| title | source · zivahub uct ac za | connector:zivahub_uct_ac_za@1.0.0 | /metadata/dc/title |