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PXD026067

PXD026067 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleIdentification of Actin Filament Interactors in Giardia lamblia
DescriptionThe deep-branching protozoan parasite Giardia lamblia is the causative agent of the intestinal disease giardiasis. Consistent with its proposed evolutionary position, many pathways are minimalistic or divergent, including its actin cytoskeleton. Giardia is the only eukaryote known to lack all canonical actin-binding proteins. Previously, our lab identified a number of non-canonical Giardia lamblia actin (GlActin) interactors; however, these proteins appeared to interact only with monomeric or globular actin (G-actin), rather than filamentous actin (F-actin). To identify interactors, we used a chemical crosslinker to preserve native interactions, followed by an anti-GlActin antibody, Protein A affinity chromatography, and liquid chromatography coupled to mass spectrometry. We found 46 putative actin interactors enriched in the conditions favoring F-actin. None of the proteins identified contain known actin-interacting motifs, and many lacked conserved domains. Each potential interactor was then tagged with the fluorescent protein mNeonGreen and visualized in live cells. We categorized the proteins based on their primary localization; localizations included ventral disc, marginal plate, nuclei, flagella, plasma membrane, and internal membranes. One protein from each category was co-localized with GlActin using immunofluorescence microscopy. We also co-immunoprecipitated one protein from each category and confirmed three interactions. Most of the localization patterns are consistent with previously demonstrated GlActin functions, but the ventral disc represents a new category of actin interactor localization. These results suggest a role for GlActin in ventral disc function, which has previously been controversial.
HostingRepositoryPRIDE
AnnounceDate2021-07-10
AnnouncementXMLSubmission_2021-07-10_13:34:08.731.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterRichard Johnson
SpeciesList scientific name: Giardia lamblia ATCC 50803; NCBI TaxID: 184922;
ModificationListNo PTMs are included in the dataset
InstrumentLTQ
Dataset History
RevisionDatetimeStatusChangeLog Entry
02021-05-18 01:43:33ID requested
12021-07-10 13:34:09announced
Publication List
Dataset with its publication pending
Keyword List
submitter keyword: Giardia, actin,
Contact List
Alexander R. Paredez
contact affiliationDepartment of Biology, University of Washington
contact emailaparedez@uw.edu
lab head
Richard Johnson
contact affiliationUniversity of Washington
contact emailrj8@uw.edu
dataset submitter
Full Dataset Link List
Dataset FTP location
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