Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Haloacetic Acids and Haloacetamides Attack Distinct Cellular Proteome Thiols

ABSTRACT: Disinfection by-products (DBPs) exposure has been linked to multiple adverse health outcomes. However, the molecular initiating events by which DBPs induce their toxicities remains unclear. Herein we combined reporter cell lines and activity-based protein profiling (ABPP) chemical proteomics to identify protein targets of six haloacetic acids (HAAs) and haloacetamides (HAMs), on the proteome-wide level. While HAAs and HAMs had similar cytotoxicities, when compared to controls, the later has 12.5 times greater Nrf2-mediated oxidative stress response, demonstrating their distinct toxicity pathways. ABPP on crude cell lysates suggested that nonspecific proteome thiol reactivity correlates with cytotoxicity. Interestingly, live cell ABPP results revealed class-specific proteins attacked by HAMs or HAAs. Subsequent proteomics analysis identified >100 unique targets per DBP. HAMs showed preferential reactivity towards disulfide oxidoreductase enzymes, accounting for their stronger Nrf2 responses. To further probe alkylation mechanisms, we directly monitored protein adducts and identified 120 and 37 unique peptides with IAM and IAA adducts, respectively. Of the later we confirmed glyceraldhye-3-phosphate dehydrogenase (GAPDH) as a key target of IAA, specifically attacked at the catalytic Cys 152. This is the first study reporting the distinct cellular protein targets of HAAs and HAMs on the proteome-wide level, which highlights their different toxicity pathways despite their similar structures.

INSTRUMENT(S): Q Exactive HF-X

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

SUBMITTER: David Hall

LAB HEAD: Hui Peng

PROVIDER: PXD018825 | Pride | 2020-11-11

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	B1.raw	Raw
	B1_2.raw	Raw
	B2.raw	Raw
	B2_2.raw	Raw
	B3.raw	Raw

Items per page:

1 - 5 of 35

Similar Datasets

Multi-Tiered Chemical Proteomic Maps of Tryptoline Acrylamide-Protein Interactions in Cancer Cells

Project description:Covalent chemistry is a versatile approach for expanding the ligandability of the human proteome. Activity-based protein profiling (ABPP) can infer the specific residues modified by electrophilic compounds through competition with broadly reactive probes. Nonetheless, the extent to which such residue-directed ABPP platforms are capable of fully mapping the protein targets of electrophilic compounds in human cells remains unclear. Here, we introduce a complementary approach that directly identifies proteins showing stereoselective reactivity with focused libraries of stereochemically-defined, alkynylated electrophilic compounds. Integration of protein- and cysteine-directed ABPP data from compound-treated human cancer cells revealed generally well-correlated target maps and highlighted specific features, such as protein size and the proteotypicity of cysteine-containing peptides, that help to explain gaps in each ABPP platform. The integrated ABPP strategy furnished stereoselective, high-engagement covalent ligands for > 300 structurally and functionally diverse human proteins, including compounds that modulate enzymes by targeting isotype-restricted and non-catalytic cysteines.

2024-06-07 | PXD042541 | Pride

Suppression of NRF2-dependent cancer growth by a covalent allosteric molecular glue

Project description:The NRF2 transcription factor is constitutively active in cancer where it functions to maintain oxidative homeostasis and reprogram cellular metabolism. NRF2-active tumors exhibit NRF2-dependency and resistance to chemo/radiotherapy. Here we developed VVD-065, a first-in-class NRF2 inhibitor that acts via an unprecedented allosteric molecular glue mechanism. In the absence of stress or mutation, NRF2 is rapidly degraded by the KEAP1-CUL3 ubiquitin-ligase complex. VVD-065 specifically and covalently adducts C151 on KEAP1, which in turn promotes KEAP1-CUL3 complex formation, leading to dramatic enhancement of NRF2 degradation. Previously reported C151-directed compounds decrease KEAP1-CUL3 interactions and stabilize NRF2, thus establishing KEAP1_C151 as a tunable regulator of the KEAP1-CUL3 complex and NRF2 stability. VVD-065 inhibited NRF2-dependent tumor growth and sensitized cancers to chemo/radiotherapy, supporting an open Phase I clinical trial (NCT05954312).

2025-01-03 | GSE278482 | GEO

IsoTOP-ABPP in Jurkat cells for cysteine reactivity profiling

Project description:Cysteine reactivity in Jurkat cell lines were assessed using IsoTOP-ABPP

2021-02-04 | PXD022151 | Pride

RNA-seq of microglia and monocyte-derived cells in Nrf2 knockout and wild-type mice after induced intracerebral hemorrhage or sham treatment

Project description:Secondary injury causes death and dependence after spontaneous intracerebral haemorrhage (ICH). Having found that ICH is associated with activation of genes regulated by the transcription factor Nrf2, particularly in mononuclear myeloid cells (for example, monocyte-derived cells (MdC) and microglia), we sought to determine the importance of Nrf2 to mononuclear myeloid cell responses and their impact on ICH pathology. We used intrastriatal injection of collagenase to induce ICH in both wild-type mice, and knockout mice with Cx3cr1-Cre-mediated excision of Nrf2 (Nfe2l2) exon 5.

2024-08-13 | E-MTAB-14324 | biostudies-arrayexpress

NAIA for cysteine profiling in HepG2 cells, protein target identification of covalent ligand and studying cysteine oxidative modifications

Project description:New cysteine probes, N-acryloylindole-alkynes (NAIAs), have been developed. We have applied NAIA-5 for cysteine profiling in HepG2 cell lysates, and the results have been compared to the conventional cysteine probe iodoacetamide-alkyne (IAA). NAIA-5 allows identification of more ligandable Cys than IAA in our experiment, and more interesting, a unique pool of Cys which have not been profiled previously by any reported cysteine probe. We have further utilized NAIA-5 for covalent ligand screening experiment. This allows us to discover hit compounds targeting the newly identified cysteines and proteins. One of the hit compounds, CL1, demonstrated good bioactivity by arresting cell cycle at G1 phase through targeting proteins associated with cell cycle and regulations. In addition, NAIA-5 has been successfully coupled with NAI compound for studying cysteine oxidative modifications in HepG2 cells by MS experiments.

2023-06-14 | PXD041264 | Pride

Reimagining high-throughput profiling of reactive cysteines for cell-based screening of large electrophile libraries

Project description:Current methods for measuring amino-acid side-chain reactivity lack the throughput needed to screen large chemical libraries for interactions across the proteome. Here we redesigned the workflow for activity-based protein profiling of reactive cysteine residues by using a smaller desthiobiotin-based probe, sample multiplexing, reduced protein starting amounts, and software to boost data acquisition in real-time on the mass spectrometer. Our method, Streamlined Cysteine Activity-Based Protein Profiling (SLC-ABPP), achieved a 42-fold improvement in sample throughput, corresponding to profiling library members at a depth of >8,000 reactive cysteine sites in 18 min per compound. We applied it to identifying the proteome-wide targets of a covalent inhibitor of mutant KRASG12C and of ibrutinib, a covalent inhibitor of BTK. In addition, we created a resource of cysteine reactivity to 285 electrophiles in three human cell lines , which includes >20,000 cysteines in >6,000 proteins per cell line. The goal of proteome-wide profiling of cysteine reactivity across thousand-member libraries under several cellular contexts is now within reach.

2021-01-26 | PXD022511 | Pride

Indolin-2-one nitroimidazole antibiotics exhibit an unexpected dual mode of action

Project description:Nitroimidazoles such as metronidazole are used as anti-infective drugs against anaerobic bacteria. Upon in vivo reduction of the nitro group, reactive radicals damage DNA and proteins in the absence of oxygen. Unexpectedly, a recent study of nitroimidazoles linked to an indolin-2-one substituent revealed potent activities against aerobic bacteria. This suggests a different, yet undiscovered mode of action (MoA). To decipher this MoA, we first performed whole proteome analysis of compound treated cells, revealing an upregulation of bacteriophage-associated proteins, indicative of DNA damage. Since DNA binding of the compound was not observed, we applied activity-based protein profiling (ABPP) for direct target discovery. Labeling studies revealed topoisomerase IV, an essential enzyme for DNA replication, as the most enriched hit in pathogenic Staphylococcus aureus cells. Subsequent topoisomerase assays confirmed the inhibition of DNA decatenation in the presence of indolin-2-one nitroimidazole with an activity comparable to ciprofloxacin, a known inhibitor of this enzyme. Furthermore, we determined significantly increased redox potentials of indolin-2-one nitroimidazoles compared to classic 5-nitroimidazoles such as metronidazole, which facilitates in vivo reduction. Overall, this study unravels that indolin-2-one functionalized nitroimidazoles feature an unexpected dual MoA, which impairs resistance development. Given the clinical application of this compound class, the new mechanism could be a starting point to mitigate resistance.

2022-11-09 | PXD033128 | Pride

Transcription profiling of mouse Keap1-null (CKO) to investigate genetic and pharmacologic activation of Nrf2 signaling

Project description:Genetic versus chemoprotective activation of Nrf2 signaling: overlapping yet distinct hepatic gene expression profiles between Keap1 knockout and triterpenoid treated mice; Loss of Nrf2 signaling increases susceptibility to acute toxicity, inflammation, and carcinogenesis in mice due to the inability to mount adaptive responses. By contrast, disruption of Keap1 (a cytoplasmic modifier of Nrf2 turnover) protects against these stresses in mice; although dominant negative mutations in Keap1 have been identified recently in some human cancers. Global characterization of Nrf2 activation is important to exploit this pathway for chemoprevention in healthy, yet at-risk individuals and also to elucidate the consequences of hijacking the pathway in Keap1-mutant human cancers. This analysis also enables a global characterization of the pharmacodynamic action of CDDO-Im at a low dose that is relevant to chemoprevention. Experiment Overall Design: Liver-targeted conditional Keap1-null (CKO) mice provide a model of genetic activation of Nrf2 signaling. By coupling global gene expression analysis of CKO mice with analysis of pharmacologic activation using the synthetic oleanane triterpenoid CDDO-Im, we are able to gain insight into pathways affected by Nrf2 activation. CDDO-Im is an extremely potent activator of Nrf2 signaling. CKO mice were used to identify genes modulated by genetic activation of Nrf2 signaling. The CKO response was compared to hepatic global gene expression changes in wild-type mice treated with CDDO-Im at a maximal Nrf2 activating dose. n=3/group, male 9 week old mice were used. Mice were treated with a single dose of vehicle (10% Cremophor-EL, 10% DMSO, and PBS) or 30 umol CDDO-Im/kg body weight by gavage and sacrificed 6 h later.

2009-05-11 | E-GEOD-15633 | biostudies-arrayexpress

RNA-seq of triple cultures of neurons, astrocytes and microglia treated with blood clot conditioned media, lipopolysaccharide, and Nrf2-activating drug CDDO-TFEA.

Project description:Secondary injury causes death and dependence after spontaneous intracerebral haemorrhage (ICH). We found that myelomononuclear Nrf2 is active and protective after ICH in mice. To investigate the roles of myelomononuclear Nrf2 in modulating cell autonomous and non-cell autonomous responses to the oxidative and inflammatory consequences of ICH we developed an in vitro system of co-cultures of primary mouse microglia, human astrocytes and rat neurons and performed RNA-seq, subsequently performing in-silico separation of read data into separate species, and thus separate cell types. Cultures were treated with blood clot condition media (CCM), lipopolysaccharide (LPS), and Nrf2-activating drug CDDO-TFEA, and experiments were performed using wild-type mouse microglia, and microglia from mice with global Nrf2 deletion.

2024-08-13 | E-MTAB-14327 | biostudies-arrayexpress

Comparison of transcriptome profile between wild-type and dao mutant plants

Project description:We identified and characterized that the rice Dioxygenase for Auxin Oxidation (dao) mutant displays pleiotropic phenotypes, including indehiscent anthers, sterile pollens, and increased free IAA levels in anthers.We conclude that DAO-mediated auxin catabolism is essential for auxin homeostasis and later stages of plant reproductive development, including anther dehiscence, and pollen viability. We used microarrays to identify differentially expressed genes in dao.

2012-09-25 | GSE41097 | GEO

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data