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Immunoprecipitation of mitochondrial ribosome complex of Toxoplasma gondii

ABSTRACT: Mitochondrial ribosomes in Apicomplexan parasite Toxoplasma gondii are expected to be divergent from all the other known mitoribosomes owing to the extremely fragmented and reduced rRNAs. To elucidate the structure and composition of these mitoribosomes, we performed immunoprecipitation of the FLAG epitope tagged bL12m subunit of mitoribosome using anti-FLAG M2 agarose beads. The elute was used to freeze CryoEM grids for single particle analysis and bound beads were subjected to Liquid chromatography–mass spectrometry (LC–MS) to analyse the co-immunoprecipitated proteins.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Toxoplasma Gondii

TISSUE(S): Permanent Cell Line Cell

DISEASE(S): Toxoplasmosis

SUBMITTER: Shikha Shikha

LAB HEAD: Lilach Sheiner

PROVIDER: PXD053944 | Pride | 2025-04-21

REPOSITORIES: Pride

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			Action	DRS
	bL12m_FLAG_IP.raw	Raw
	bL12m_FLAG_IP_searchfile.txt	Txt
	checksum.txt	Txt

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Publications

Numerous rRNA molecules form the apicomplexan mitoribosome via repurposed protein and RNA elements.

Shikha Shikha S Tobiasson Victor V Ferreira Silva Mariana M Ovciarikova Jana J Beraldi Dario D Mühleip Alexander A Sheiner Lilach L

Nature communications 20250118 1

Mitochondrial ribosomes (mitoribosomes) are essential, and their function of synthesising mitochondrial proteins is universal. The core of almost all mitoribosomes is formed from a small number of long and self-folding rRNA molecules. In contrast, the mitoribosome of the apicomplexan parasite Toxoplasma gondii assembles from over 50 extremely short rRNA molecules. Here, we use cryo-EM to discover the features that enable this unusual mitoribosome to perform its function. We reveal that poly-A ta ...[more]

PMID: 39827269

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