PXD026067-1

PXD026067 is an original dataset announced via ProteomeXchange.

Title	Identification of Actin Filament Interactors in Giardia lamblia
Description	The 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.
HostingRepository	PRIDE
AnnounceDate	2021-07-10
AnnouncementXML	Submission_2021-07-10_13:34:08.731.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Richard Johnson
SpeciesList	scientific name: Giardia lamblia ATCC 50803; NCBI TaxID: 184922;
ModificationList	No PTMs are included in the dataset
Instrument	LTQ

Revision	Datetime	Status	ChangeLog Entry
0	2021-05-18 01:43:33	ID requested
⏵ 1	2021-07-10 13:34:09	announced

Dataset with its publication pending

submitter keyword: Giardia, actin,

Alexander R. Paredez
contact affiliation	Department of Biology, University of Washington
contact email	aparedez@uw.edu
lab head
Richard Johnson
contact affiliation	University of Washington
contact email	rj8@uw.edu
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2021/07/PXD026067

PRIDE project URI

• PRIDE
  1. PXD026067
     1. Label: PRIDE project
     2. Name: Identification of Actin Filament Interactors in Giardia lamblia