Proteome-wide crosslinking mass spectrometry reveals novel components of essential complexes in Toxoplasma
- 1Whitehead Institute for Biomedical Research, Cambridge, MA, U.S.A.; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, U.S.A.
Abstract
Protein-protein interactions underpin nearly all cellular processes, yet systematic definition of these networks remains limited outside a few model organisms. As a result, the architectures of essential complexes in many divergent lineages remain poorly characterized. Here we developed a high-coverage crosslinking mass spectrometry framework to map the proteome-wide interactome of the model apicomplexan parasite Toxoplasma gondii. From 29,624 crosslinked peptide pairs, we resolved a network of 2,859 protein-protein interactions that we integrated with structural modeling to resolve interaction interfaces. We identified and validated previously unrecognized components of essential protein complexes, including a structurally distinct ATP synthase subcomplex containing a highly divergent, apicomplexan-specific subunit essential for parasite fitness. Beyond revealing unexpected diversification of core mitochondrial machinery, these findings provide a general strategy to define the molecular architecture of divergent organisms and represent a foundational resource for hypothesis generation, structural inference, and discovery of lineage-specific vulnerabilities in pathogen biology.
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BibTeX
@misc{butterworth2026proteomewide,
title = {{Proteome-wide crosslinking mass spectrometry reveals novel components of essential complexes in Toxoplasma}},
author = {S. Butterworth and A. L. Gin and S. Shikha and I. Tengganu and J. Rush and T. Duraisingh and V. Sodeinde and L. Lemgruber and F. Schulte and K. Hu and L. Sheiner and S. Ovchinnikov and S. Lourido},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.01.748362},
url = {https://www.biorxiv.org/content/10.64898/2026.09.01.748362v1},
note = {CC BY-NC},
}
RIS
TY - UNPB
TI - Proteome-wide crosslinking mass spectrometry reveals novel components of essential complexes in Toxoplasma
AU - S. Butterworth
AU - A. L. Gin
AU - S. Shikha
AU - I. Tengganu
AU - J. Rush
AU - T. Duraisingh
AU - V. Sodeinde
AU - L. Lemgruber
AU - F. Schulte
AU - K. Hu
AU - L. Sheiner
AU - S. Ovchinnikov
AU - S. Lourido
PY - 2026
DA - 2026/09/01
PB - bioRxiv
DO - 10.64898/2026.09.01.748362
UR - https://www.biorxiv.org/content/10.64898/2026.09.01.748362v1
AB - Protein-protein interactions underpin nearly all cellular processes, yet systematic definition of these networks remains limited outside a few model organisms. As a result, the architectures of essential complexes in many divergent lineages remain poorly characterized. Here we developed a high-coverage crosslinking mass spectrometry framework to map the proteome-wide interactome of the model apicomplexan parasite Toxoplasma gondii. From 29,624 crosslinked peptide pairs, we resolved a network of 2,859 protein-protein interactions that we integrated with structural modeling to resolve interaction interfaces. We identified and validated previously unrecognized components of essential protein complexes, including a structurally distinct ATP synthase subcomplex containing a highly divergent, apicomplexan-specific subunit essential for parasite fitness. Beyond revealing unexpected diversification of core mitochondrial machinery, these findings provide a general strategy to define the molecular architecture of divergent organisms and represent a foundational resource for hypothesis generation, structural inference, and discovery of lineage-specific vulnerabilities in pathogen biology.
ER -