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Quantitative Interactomics of Lck-TurboID in Living Human T Cells Unveils T Cell Receptor Stimulation-Induced Proximal Lck Interactors
While Lck has been widely recognized to play a pivotal role in the initiation of the T cell receptor (TCR) signaling pathway, an understanding of the precise regulation of Lck in T cells upon TCR activation remains elusive. Investigation of protein-protein interaction (PPI) using proximity labeling …
In vivo interactome profiling by enzyme‐catalyzed proximity labeling
Enzyme-catalyzed proximity labeling (PL) combined with mass spectrometry (MS) has emerged as a revolutionary approach to reveal the protein-protein interaction networks, dissect complex biological processes, and characterize the subcellular proteome in a more physiological setting than before. The enzymatic tags are being upgraded to improve …
AirID, a novel proximity biotinylation enzyme, for analysis of …
Figure 4—source data 1: Cell growth analysis data relating to Figure 4E.This excel file contains cell growth analysis data obtained using the MTS assay shown in Figure 4E.Main signal (490 nm) and background signal (650 nm) were detected. The graph in Figure 4E was generated using a ratio calculated on the basis of value subtracted background signal value.
TurboID functions as an efficient biotin ligase for BioID …
To evaluate the efficiency of TurboID relative to BioID2, we analyzed samples from embryos expressing TurboID or BioID2 and supplemented their culture media with 0, 100, or 500 μM biotin. As before, the mRNA constructs were injected into Xenopus two-cell embryos and allowed to develop to the blastula stage (stage 7). ...
Efficient TurboID-based proximity labelling method for identifying …
TurboID, a proximity labelling method based on mutant biotin ligase, is an efficient new technique for recognizing protein-protein interactions and has been successfully applied to living cells. Sialic acid is typically the terminal monosaccharide attached to many glycoproteins and plays many import …
When less is more
In recent years, proximity labelling has established itself as an unbiased and powerful approach to map the interactome of specific proteins. Generally, protein fusions with labelling enzymes are transiently overexpressed to perform these experiments. Using a pipeline for the rapid generation CRISPR-Cas9 knock-ins (KIs) based on antibiotic …
TurboID Screening of the OmpP2 Protein Reveals Host Proteins …
Expression of His-TurboID-OmpP2 and His-TurboID and validation of their biotinylation efficiencies. To produce a fusion protein for proximal biotin labeling, the OmpP2 gene of G. parasuis serotype 5 was fused to the TurboID gene with a 6-amino-acid linker (GGSGGS) in the expression vector pET-28a and a His tag fused to the 5′ end of TurboID (Fig. 1A).
RETICULON-LIKE PROTEIN B2 is a proviral factor co-opted for the …
Endomembrane remodeling to form a viral replication complex (VRC) is crucial for a virus to establish infection in a host. Although the composition and function of VRCs have been intensively studied, host factors involved in the assembly of VRCs for plant RNA viruses have not been fully explored. Tu …
Potential application of TurboID-based proximity labeling in …
Introduction Plant life needs certain natural factors such as temperature, moisture and nutrition, while plants often suffer from environmental stresses during development including changes of temperature, salinity, water, light, nutrient availability, and toxic chemicals (Vanstraelen and Benkova, 2012; Skalak et al., 2021; Zhou et al., 2021).
Split-TurboID enables contact-dependent proximity labeling in …
Browse Alice Ting Cho et al Split-TurboID enables contact-dependent proximity labeling in cells. Cho KF, Branon TC, Rajeev S, Svinkina T, Udeshi ND, Thoudam T, Kwak C, Rhee HW, Lee IK, Carr SA, Ting AY Proc Natl Acad Sci U S A. 2020 May 18. pii
16 Best Things To Do in Tbilisi, Georgia (3-Day Guide)
Tbilisi in Georgia is all about great food, wine, architecture, and picturesque corners. Here are tips for the places to visit in Tbilisi. 3. Gallery 27 Gallery 27 is an eye-catching house with impressive carvings and has a photogenic staircase entrance decorated with ...
TurboID-mediated surface protein biotinylation to inhibit the …
Staphylococcus aureus (S. aureus) infection is major cause of nosocomial infections. Antibiotic treatment for S. aureus remains the primary solution for managing S. aureus infections, which, however, increases the risk of antibiotic resistance. To broaden the resolutions on S. aureus infection, here we report TurboID-mediated protein proximity …
Mapping the signaling network of BIN2 kinase using TurboID …
Combining TurboID-mediated proximity labeling with quantitative phosphoproteomics identifies BIN2 signaling components including kinase substrates in vivo, Abstract Elucidating enzyme–substrate relationships in posttranslational modification (PTM) networks is ...
TurboID-EV: Proteomic Mapping of Recipient Cellular Proteins …
Overview of the TurboID-EV System Among currently available biotin ligases and peroxidases, we chose TurboID because it has a higher biotinylation activity and higher temporal resolution (approximately >10 min) than other biotin ligases and does not require H 2 O 2 treatment for biotinylation in contrast to peroxidases. 15 We chose HEK293T as a …
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