Dataset Information

Dynamic T. vaginalis secretome and secretion of β-amalyses

ABSTRACT: During infection of the urogenital tract, the parasite Trichomonas vaginalis is challenged by factors such as nutrient limitation, the host immune response and coexistence with other members of the microbial fauna. Thus establishment of trichomonad infection depends on multifactorial host-parasite interactions, that involve both contact-dependent mechanisms, such as adherence to vaginal epithelial cells, contact-dependent extracellular killing of host cells, and active phagocytosis of host cells and bacteria, as well as contact independent mechanisms that include secretion of soluble biologically active molecules. To get more insight into the secreted proteome of T. vaginalis we detected and quantified released proteins at consecutive time points. Out of 2072 detected proteins, 89 were estimated to significantly increase in time. These include leishmaniolysin-like metallopeptidases, cathepsin-like cystein peptidases and a number of proteins involved in sugar metabolism, most notably beta-amylases, which may be significant in utilization of host-cell derived glycogen. We further discovered a large group of secreted hypothetical proteins with predicted triple tyrosine domain that may facilitate interaction with the extracellular matrix of the host epithelium.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Trichomonas Vaginalis

TISSUE(S): Cell Suspension Culture, Cell

DISEASE(S): Trichomoniasis

SUBMITTER: Karel Harant

LAB HEAD: Jan Tachezy

PROVIDER: PXD007034 | Pride | 2017-12-19

REPOSITORIES: pride

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Publications

Dynamic secretome of Trichomonas vaginalis: Case study of β-amylases.

Štáfková Jitka J Rada Petr P Meloni Dionigia D Žárský Vojtěch V Smutná Tamara T Zimmann Nadine N Harant Karel K Pompach Petr P Hrdý Ivan I Tachezy Jan J

Molecular & cellular proteomics : MCP 20171212 2

The secretion of virulence factors by parasitic protists into the host environment plays a fundamental role in multifactorial host-parasite interactions. Several effector proteins are known to be secreted by Trichomonas vaginalis, a human parasite of the urogenital tract. However, a comprehensive profiling of the T. vaginalis secretome remains elusive, as do the mechanisms of protein secretion. In this study, we used high-resolution label-free quantitative MS to analyze the T. v ...[more]

PMID: 29233912

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Project description:Lysosomes represent a central degradative compartment of eukaryotes, yet little is known about 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 was reported in other eukaryotes. In this study, we investigated the phagolysosomal proteome of the human parasite Trichomonas vaginalis, protein targeting and 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 phagocytosis of lactoferrin-covered magnetic nanoparticles. The analysis resulted in the lysosomal proteome of 462 proteins, which were sorted into 21 functional classes. Hydrolases represent the largest functional class and include proteases, lipases, phosphatases, glycosidases, and other hydrolases. Identification of a large set of proteins involved in vesicular trafficking (80) and turnover of actin cytoskeleton rearrangement (29) indicate high dynamics of phagolysosomal compartment. Several lysosomal proteins including the cysteine protease TvCP2 were previously shown to be secreted. Our experiment showed that secretion of TvCP2 was strongly inhibited by chloroquine that increased intralysosomal pH and thus indicated TvCP2 secretion through lysosomes rather than the classical secretory pathway. Unexpectedly, we identified in the phagolysosomal proteome divergent homologs of M6P receptor TvMPR, 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 that possesses two glycosylation sites. Mutation of any of the sites redirected CLCP to the secretory pathway. Similarly, 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 recognition marker for lysosomal hydrolases. Whether TvMPR or other possible receptors are involved in this process and what is the precise structure of the lysosomal recognition marker need to be clarified in future studies.

Project 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.

			Action	DRS
	20170222_Tachac_search_revize.zip	Other
	Pompo_MS.7z.001	Other
	Pompo_MS.7z.002	Other
	max_quant_search.zip	Other
	mqpar.xml	Xml

Dataset Information

Dynamic T. vaginalis secretome and secretion of β-amalyses

Dataset's files

Publications

Dynamic secretome of <i>Trichomonas vaginalis</i>: Case study of β-amylases.

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