Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

TEAD1-DBD-DNA interaction studied by structural MS

ABSTRACT: TEAD transcription factors mediate gene expression regulation through interactions with their DNA response M-CAT motif. They are active mainly during growth and development and induce gene expression of proteins involved in cell proliferation, differentiation or apoptosis prevention. TEADs and many of their target proteins are known to be upregulated in several types of cancers. To study the interaction of DNA binding domain of TEAD1 with M-CAT motifs of different orientations originating from regulatory regions of human genes (CTGF, SRF, C-MYC and GLUT1), dissociation constant of each complex was determined using fluorescence anisotropy assay. Structural mass spectrometry techniques (H/D exchange and quantitative chemical cross-linking) and molecular docking were utilized for structural characterization of the binding interface. In-vitro results were correlated with situation in living cells studied by ChIP and Q-PCR.

INSTRUMENT(S): Bruker Daltonics solarix series

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Jurkat Cell

SUBMITTER: Ruzena Filandrova

LAB HEAD: Petr Novak

PROVIDER: PXD012127 | Pride | 2020-12-08

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Similar Datasets

Structural proteomics based investigation of DNA binding domain (DBD) of FOXO4 transcription factor in complex with DNA binding element (DNA)

Project description:The limited information available on the structure of complexes involving transcription factors and cognate DNA response elements represents a major obstacle in the quest to understand their mechanism of action at the molecular level. We implemented a concerted structural proteomics approach, which combined hydrogen-deuterium exchange (HDX), quantitative protein-protein and protein-nucleic acid cross-linking (XL), and homology analysis, to model the structure of the complex between the full-length DNA binding domain (DBD) of FOXO4 and its DNA binding element (DBE).

2020-05-26 | PXD013969 | Pride

Influence of cross-linker polarity on selectivity towards lysine side chains

Project description:Even though the amine reactive BS2G and DSG cross-linkers have the same length of spacer and are based on N-hydroxysuccinimidic group, our data showed that each of them formed preferentially different cross-links. We demonstrated that the choice of cross-linker can have a significant impact on the output data for structural characterization of biomolecules. Using equimolar mixtures of DSG with d6-BS2G, and BS2G with d6-DSG, we established that the polar BS2G preferentially bound to polar regions of modified molecule, whereas non-polar DSG bound to hydrophobic regions. This phenomenon established that the mixture of polar and non-polar cross-linkers acted as an efficient tool for the determination of distance constraints in proteins.

2020-02-28 | PXD017299 | Pride

EPIC-XS H2020 alpha synuclein IMIC

Project description:Hydrogen/deuterium exchange and quantitative chemical cross-linking of monomeric and fibrilar alpha synuclein

2020-08-03 | PXD019659 | Pride

Epitope and Paratope Mapping of TNFα/Infliximab complex by cross-linking mass spectrometry and integrative modeling reveals high level of interaction specificity

Project description:Mass spectrometry (MS) has become one of the core technologies to study protein structures, protein-ligand and protein-protein interactions. Chemical cross-linking combined with mass spectrometry (XL-MS) is quickly spreading over different biochemistry laboratories for the continuous increase of its biological applications and standardized protocols that make it attractive for academia and industry research scientists. In recent years, XL-MS has developed as a powerful technique in structural biology, from studying structures of purified proteins in vitro to deciphering intricate protein-protein interaction (PPI) networks in cells, organelles and tissues on a system-wide scale. However, the number of XL-MS studies for the inves-tigation of epitope/paratope mapping of antigen-antibody complexes is still limited up to now. In this manuscript, we devel-oped a simple, fast and automated workflow to define the interaction interface between the chimeric antibody Infliximab (IFX) and its own target, the Tumor Necrosis Factor alpha (TNFα). This workflow integrates XL-MS data to identify areas in proximity to the binding regions and epitope/paratope probability calculation to pinpoint the antigen-antibody binding sites. The data are combined to generate refined 3D models of the antigen-antibody complex which closely resemble the X-ray IFX/TNFα structure and capture their correct interaction surface. The present workflow can be applied to investigate any antigen-antibody complex, even when no previous structural knowledge is available. This strategy is a fascinating oppor-tunity to thoroughly characterize antigen-antibody interactions and to allow the understanding of their binding mechanisms in order to properly design better antibody therapeutics in the future

2024-02-27 | PXD043347 | Pride

CL-MS OF hE1a (DHTKD1) or G729R hE1a with hE2o

Project description:CL-MS OF hE1a (DHTKD1) or G729R hE1a with hE2o to identify the possible difference in the interactions between hE1a-hE2o and G729R hE1a-hE2o

2020-04-22 | PXD017792 | Pride

Structural basis for the specificity of PPM1H phosphatase for Rab GTPases

Project description:LRRK2 serine/threonine kinase is associated with inherited Parkinson’s disease. LRRK2 phosphorylates a subset of Rab GTPases within their switch 2 motif to control their interactions with effectors. Recent work has shown that the metal-dependent protein phosphatase PPM1H counteracts LRRK2 by dephosphorylating Rabs. PPM1H is highly selective and closely related PPM1J/M exhibit virtually no activity toward substrates such as Rab8a phosphorylated at T72 (pT72). Here we have identified the structural determinant of PPM1H specificity for Rabs. The crystal structure of PPM1H reveals a conserved catalytic domain that adopts a β-sandwich fold. The striking difference is that PPM1H has evolved a 110-residue flap domain that punctuates the catalytic domain. The flap domain distantly resembles tudor domains that interact with histones in the context of epigenetics. Cellular assays and 3-D modelling suggest that the flap domain encodes the docking motif for phosphorylated Rabs. Consistent with this hypothesis, a PPM1J chimera with the PPM1H flap domain dephosphorylates pT72 of Rab8a with a higher specific activity than PPM1H. Therefore, PPM1H has acquired a Rab-specific interaction domain within a conserved phosphatase fold.

2021-08-11 | PXD026367 | Pride

Quality control at the cell surface prevents accumulation of damaged L-type CaV1.2 calcium channels

Project description:Defective ion channel turnover and clearance of damaged proteins are associated with aging and neurodegeneration. The L-type CaV1.2 voltage-gated calcium channel mediate depolarization-induced calcium signals in heart and brain. Here, we determined the interaction surface between the L-type calcium channel CaVβ subunit and actin using cross-linking mass spectrometry and protein-protein docking, and uncovered a role in replenishing damaged CaV1.2 channels. Computational and in vitro mutagenesis identified hotspots in CaVβ that decrease its affinity for actin but not for CaV1.2. Coexpression of an actin-association-deficient CaVβ mutant with the CaV1.2 channel downregulated current amplitudes with a concomitant reduction in the number of functionally available channels. Neither alterations in the single-channel properties nor changes in the total number of channels at the cell surface were found, indicating that current inhibition resulted from a build-up of conduction-defective channels. Our findings established CaVβ–actin interaction as a key player for selective monitoring and clearing corrupted CaV proteins to ensure the maintenance of a functional pool of channels and proper calcium signal transduction. The CaVβ–actin molecular model introduces a potentially druggable protein-protein interface to intervene CaV-mediated signaling processes.

2024-02-16 | PXD044723 | Pride

Quality control at the cell surface prevents accumulation of damaged L-type CaV1.2 calcium channels

2024-02-16 | PXD044630 | Pride

Cross-linking of mouse laminin 111

Project description:Laminin, a ~800 kD heterotrimeric protein, is a major functional component of the extracellular matrix, contributing to tissue development and maintenance. We used cross-linking and mass spectrometry to determine the order of subunits in the ~750 Å trimeric coiled-coil oligomerization domain of laminin and to achieve an estimate of the register of the three subunits along the length of the coiled coil.

2016-12-22 | PXD004898 | Pride

Enabling Photoactivated Cross-linking Mass Spectrometric Analysis of Protein Complexes by Novel MS-cleavable Cross-linkers

Project description:Cross-linking mass spectrometry (XL-MS) is a powerful tool for studying protein-protein interactions and elucidating architectures of protein complexes. While residue-specific XL-MS studies have been very successful, accessibility of interaction regions non-targetable by specific chemistries remain difficult. Photochemistry has shown great potential in capturing those regions due to nonspecific reactivity, but low yields and high complexities of photocross-linked products have hindered their identification, limiting current studies predominantly to single proteins. Here, we describe the development of three novel MS-cleavable heterobifunctional cross-linkers, namely SDASO (Succinimidyl diazirine sulfoxide), to enable fast and accurate identification of photocross-linked peptides by MSn. The MSn-based workflow allowed SDASO XL-MS analysis of the yeast 26S proteasome, demonstrating the feasibility of photocross-linking of large protein complexes for the first time. Comparative analyses have revealed that SDASO cross-linking is robust and captures interactions complementary to residue-specific reagents, providing the foundation for future applications of photocross-linking in complex XL-MS studies.

2021-05-07 | PXD022690 | Pride

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

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