PXD027121-1

PXD027121 is an original dataset announced via ProteomeXchange.

Title	In vivo structure-function analysis and redox interactomes of Leishmania tarentolae Erv
Description	Import and oxidative folding of proteins in the mitochondrial intermembrane space differ among eukaryotic lineages. While opisthokonts such as yeast rely on the receptor and oxidoreductase Mia40 in combination with the Mia40:cytochrome c oxidoreductase Erv, kinetoplastid parasites and other excavates lack Mia40 but have a functional Erv homologue. Whether excavate Erv homologues rely on a Mia40 replacement or directly interact with imported protein substrates remains controversial. Here we used the CRISPR-Cas9 system to generate a set of homozygous tagged and untagged point and deletion mutants of LTERV from the kinetoplastid model parasite Leishmania tarentolae. Modifications of the shuttle cysteine motif of LtErv were lethal, whereas replacement of clamp residue Cys17 or removal of the kinetoplastida-specific second (KISS) domain had no impact on parasite viability under standard growth conditions. However, removal of the KISS domain rendered parasites sensitive to heat stress and led to the accumulation of homodimeric and mixed LtErv disulfides. We therefore determined and compared the redox interactomes of tagged wild-type LtErv and LtErvΔKISS using SILAC and quantitative mass spectrometry. While the Mia40-replacement candidate Mic20 and all but one typical substrates with twin Cx3C- or twin Cx9C-motifs were absent in both redox interactomes, we identified a small set of alternative interaction candidates with potential redox-active cysteine residues. In summary, our study reveals parasite-specific intracellular structure-function relationships and redox interactomes of LtErv with implications for current hypotheses on mitochondrial protein import in non-opisthokonts.
HostingRepository	PRIDE
AnnounceDate	2021-07-09
AnnouncementXML	Submission_2021-07-09_02:21:22.391.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	David Zimmer
SpeciesList	scientific name: Leishmania tarentolae; NCBI TaxID: 5689;
ModificationList	acetylated residue; monohydroxylated residue
Instrument	TripleTOF 5600; Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2021-07-05 21:55:50	ID requested
⏵ 1	2021-07-09 02:21:22	announced

Dataset with its publication pending

submitter keyword: Proteomics, Leishmania tarentolae, SILAC, IP

Marcel Deponte
contact affiliation	Biochemistry, TU Kaiserslautern, Kaiserslautern, Germany
contact email	deponte@chemie.uni-kl.de
lab head
David Zimmer
contact affiliation	TU KL - CSB
contact email	davidzimmer91@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD027121
     1. Label: PRIDE project
     2. Name: In vivo structure-function analysis and redox interactomes of Leishmania tarentolae Erv