Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Recombinant trichomonas vaginalis proteasome native gel digest

ABSTRACT: Recombinant tv20s expressed in s frugiperda and purified was subject to native gel and digested with trypsin for proteomics. Data was searched against fasta file which is combined recombinant proteasome sequences and all uniprot available s. frugiperda sequences as of 06/2023.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Spodoptera Frugiperda (ncbitaxon:7108) Trichomonas Vaginalis (ncbitaxon:5722)

SUBMITTER: Samuel Myers Anthony O'Donoghue

PROVIDER: MSV000092107 | MassIVE | Mon Jun 05 13:54:00 BST 2023

SECONDARY ACCESSION(S): PXD042740

REPOSITORIES: MassIVE

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Recombinant trichomonas vaginalis proteasome native gel digest

Project description:Recombinant proteasome (expressed in S. frugiperda) was loaded to native gel. Gel band was visualized via silver stain and excised for proteomics. MS data was searched against a fasta file including the proteome of S. frugiperda and the cloned T. vaginalis proteins.

2023-05-26 | MSV000092044 | MassIVE

Native gel digest of recombinantly expressed trichomonas vaginalis proteasome

Project description:Tv20S expressed recombinantly in S frugiperda, purified, excised from native gel and digested with trypsin.

2024-04-19 | MSV000094569 | MassIVE

Structural elucidation of recombinant Trichomonas vaginalis 20S proteasome bound to covalent inhibitors.

Project description:The proteasome is a proteolytic enzyme complex essential for protein homeostasis in mammalian cells and protozoan parasites like Trichomonas vaginalis (Tv), the cause of the most common, non-viral sexually transmitted disease. Tv and other protozoan 20S proteasomes have been validated as druggable targets for antimicrobials. However, low yields and purity of the native proteasome have hindered studies of the Tv 20S proteasome (Tv20S). We address this challenge by creating a recombinant protozoan proteasome by expressing all seven α and seven β subunits of Tv20S alongside the Ump-1 chaperone in insect cells. The recombinant Tv20S displays biochemical equivalence to its native counterpart, confirmed by various assays. Notably, the marizomib (MZB) inhibits all catalytic subunits of Tv20S, while the peptide inhibitor carmaphycin-17 (CP-17) specifically targets β2 and β5. Cryo-electron microscopy (cryo-EM) unveils the structures of Tv20S bound to MZB and CP-17 at 2.8 Å. These findings explain MZB's low specificity for Tv20S compared to the human proteasome and demonstrate CP-17's higher specificity. Overall, these data provide a structure-based strategy for the development of specific Tv20S inhibitors to treat trichomoniasis.

| S-EPMC11452676 | biostudies-literature

Structural elucidation of recombinant <i>Trichomonas vaginalis</i> 20S proteasome bound to covalent inhibitors.

Project description:Proteasomes are essential for protein homeostasis in mammalian cells1-4 and in protozoan parasites such as Trichomonas vaginalis (Tv).5 Tv and other protozoan 20S proteasomes have been validated as druggable targets.6-8 However, in the case of Tv 20S proteasome (Tv20S), biochemical and structural studies were impeded by low yields and purity of the native proteasome. We successfully made recombinant Tv20S by expressing all seven α and seven β subunits together with the Ump-1 chaperone in insect cells. We isolated recombinant proteasome and showed that it was biochemically indistinguishable from the native enzyme. We confirmed that the recombinant Tv20S is inhibited by the natural product marizomib (MZB)9 and the recently developed peptide inhibitor carmaphycin-17 (CP-17)8,10. Specifically, MZB binds to the β1, β2 and β5 subunits, while CP-17 binds the β2 and β5 subunits. Next, we obtained cryo-EM structures of Tv20S in complex with these covalent inhibitors at 2.8Å resolution. The structures revealed the overall fold of the Tv20S and the binding mode of MZB and CP-17. Our work explains the low specificity of MZB and higher specificity of CP-17 towards Tv20S as compared to human proteasome and provides the platform for the development of Tv20S inhibitors for treatment of trichomoniasis.

| S-EPMC10462138 | biostudies-literature

20S Proteasome as a Drug Target in Trichomonas vaginalis.

Project description:Trichomoniasis is a sexually transmitted disease with hundreds of millions of annual cases worldwide. Approved treatment options are limited to two related nitro-heterocyclic compounds, yet resistance to these drugs is an increasing concern. New antimicrobials against the causative agent, Trichomonas vaginalis, are urgently needed. We show here that clinically approved anticancer drugs that inhibit the proteasome, a large protease complex with a critical role in degrading intracellular proteins in eukaryotes, have submicromolar activity against the parasite in vitro and on-target activity against the enriched T. vaginalis proteasome in cell-free assays. Proteomic analysis confirmed that the parasite has all seven α and seven β subunits of the eukaryotic proteasome although they have only modest sequence identities, ranging from 28 to 52%, relative to the respective human proteasome subunits. A screen of proteasome inhibitors derived from a marine natural product, carmaphycin, revealed one derivative, carmaphycin-17, with greater activity against T. vaginalis than the reference drug metronidazole, the ability to overcome metronidazole resistance, and reduced human cytotoxicity compared to that of the anticancer proteasome inhibitors. The increased selectivity of carmaphycin-17 for T. vaginalis was related to its >5-fold greater potency against the β1 and β5 catalytic subunits of the T. vaginalis proteasome than against the human proteasome subunits. In a murine model of vaginal trichomonad infection, proteasome inhibitors eliminated or significantly reduced parasite burden upon topical treatment without any apparent adverse effects. Together, these findings validate the proteasome of T. vaginalis as a therapeutic target for development of a novel class of trichomonacidal agents.

| S-EPMC6811414 | biostudies-literature

Trichomonas vaginalis 20s proteasome catalytic subunits denaturing gel proteomics

Project description:Sample 7 = top band, sample 8 = middle band, sample 9 = lowest band. Purified tv20S was incubated with 2uM MV151 probe overnight and imaged. The three bands visualized were excised and trypsin digested for proteomics. The data was searched against the T. vaginalis proteome.

2023-06-01 | MSV000092082 | MassIVE