PXD029152-1

PXD029152 is an original dataset announced via ProteomeXchange.

Title	Proteomic analysis of Trichomonas vaginalis phagolysosomes, lysosomal targeting, and unconventional secretion of cysteine peptidases
Description	Lysosome represent a central degradative compartment of eukaryote cells, yet little is known about the biogenesis and function of this organelle in parasitic protists. Whereas the mannose-6 phosphate (M6P)-dependent system is dominant for lysosomal targeting in metazoans, oligosaccharide-independent sorting has been reported in other eukaryotes. In this study, we investigated the phagolysosomal proteome of the human parasite Trichomonas vaginalis, its protein targeting and the involvement of lysosomes in hydrolase secretion. The organelles were purified using conventional Percoll and OptiPrep density gradient centrifugation and a novel purification protocol based on the phagocytosis of lactoferrin-covered magnetic nanoparticles. The analysis resulted in a lysosomal proteome of 462 proteins, which were sorted into 21 functional classes. Hydrolases represented the largest functional class and included proteases, lipases, phosphatases, and glycosidases. Identification of a large set of proteins involved in vesicular trafficking (80) and turnover of actin cytoskeleton rearrangement (29) indicate a dynamic phagolysosomal compartment. Several lysosomal proteins, including the cysteine protease TvCP2, were previously shown to be secreted. Our experiments showed that secretion of TvCP2 was strongly inhibited by chloroquine, which increases intralysosomal pH, thus indicating that TvCP2 secretion occurs through lysosomes rather than the classical secretory pathway. Unexpectedly, we identified divergent homologues of the M6P receptor TvMPR in the phagolysosomal proteome, although T. vaginalis lacks enzymes for M6P formation. To test whether oligosaccharides are involved in lysosomal targeting, we selected the lysosome-resident cysteine protease CLCP, which possesses two glycosylation sites. Mutation of any of the sites redirected CLCP to the secretory pathway. Similarly, the introduction of glycosylation sites to secreted β-amylase redirected this protein to lysosomes. Thus, unlike other parasitic protists, T. vaginalis seems to utilize glycosylation as a recognition marker for lysosomal hydrolases. Whether TvMPR or other possible receptors are involved in this process and what the precise structure of the lysosomal recognition marker needs to be clarified in future studies.
HostingRepository	PRIDE
AnnounceDate	2021-10-29
AnnouncementXML	Submission_2021-10-29_04:56:10.788.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Karel Harant
SpeciesList	scientific name: Trichomonas vaginalis; NCBI TaxID: 5722;
ModificationList	iodoacetamide derivatized residue
Instrument	Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2021-10-15 06:33:46	ID requested
⏵ 1	2021-10-29 04:56:11	announced

Dataset with its publication pending

submitter keyword: Trichomonas vaginalis, phagolysosome, proteome, cysteine peptidase, glycosylation, mannose-6-phosphate receptor

Jan Tachezy
contact affiliation	Department of Parasitology, Faculty of Science, Charles University BIOCEV, Průmyslová 595, 252 50 Vestec, Czech Republic
contact email	jan.tachezy@natur.cuni.cz
lab head
Karel Harant
contact affiliation	Charles University
contact email	harant@natur.cuni.cz
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD029152
     1. Label: PRIDE project
     2. Name: Proteomic analysis of Trichomonas vaginalis phagolysosomes, lysosomal targeting, and unconventional secretion of cysteine peptidases