Protein tyrosine (pTyr) phosphorylation is a common post-translational modification which can create novel recognition motifs for protein interactions and cellular localisation, affect protein stability, and regulate enzyme activity. (embl.de)
The bacterial SH2 domains are not only capable of binding proteins or peptides in a Tyr phosphorylation-dependent manner, some bind pTyr itself with micromolar affinities, a property not observed for mammalian SH2 domains. (rcsb.org)
He has an extensive track-record in quantitative proteomics, introducing metabolic stable isotope labeling in multicellular organisms such as Drosophila and C. elegans, using SILAC for studying stem cell and B cell differentiation, and stable isotope labeling by using chemical approaches, for example to follow differential pTyrphosphorylation in differentiating stem cells. (uu.nl)
Specificity1
The Legionella SH2 domains feature the SH2 fold and a pTyr-binding pocket, but lack a specificity pocket found in a typical mammalian SH2 domain for recognition of sequences flanking the pTyr residue. (rcsb.org)
Pocket2
The phosphate group of Tyr is tightly bound in the conserved pTyr binding pocket. (eu.org)
The N-terminal region is most conserved and contains the pTyr binding pocket. (eu.org)
Domain1
The binding affinity of an SH2 domain to a pTyr containing ligand is moderate, with the typical affinity range between 0.1 µМ to 10 µМ for equilibrium dissociation constant values (Kd) ( Kaneko,2012 ). (eu.org)
PThr3
The Cell-Based ERK1/2 (pThr 202 / pTyr 204 ) ELISA kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in cells. (sigmaaldrich.com)
It can be used for measuring the relative amount of ERK1/2 (pThr 202 / pTyr 204 ) phosphorylation and screening the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators in cultured human, mouse and rat cell lines. (sigmaaldrich.com)
Furthermore, using stable isotope labeling by amino acids in cell culture (SILAC), we were able to quantified over 400 phosphorylation sites (pTyr, pSer, and pThr) that were responsive to FLT3 inhibition in FLT3 driven human leukemia cell lines. (nih.gov)
Phosphorylation sites1
Using synthetic pTyr-containing peptides that correspond to natural phosphorylation sites in proteins, several groups have demonstrated that PTPases display a range of kcat/Km values for these relatively short peptide substrates. (watsonbio.com)
Inhibitors2
Some of her contributions to the laboratory include aiding in a genome wide CRISPR screen to gain mechanistic insights of pathway crosstalk with PI3K/Akt signaling in ovarian cancer, exploring hits from another genome wide CRISPR screen in triple negative breast cancer (TNBC) with hopes to identify druggable genetic targets that work in synergy with PI3K/Akt inhibitors, and characterizing pTyr sites on metabolic enzymes that are important in TNBC. (nih.gov)
Historically, our emphasis in this area has been on inhibitors of phosphotyrosyl (pTyr)-dependent binding interactions, which are mediated by src homology 2 (SH2) domains and on protein-tyrosine phosphatase (PTP) inhibitors. (nih.gov)
Peptides1
For this work, we designed peptidomimetics as conformationally constrained analogs of natural Grb2 SH2 domain-bound pTyr-containing peptides. (nih.gov)
Tyrosine2
Tyrosine phosphorylation (pTyr) plays a fundamental role in cellular processes and is commonly dysregulated in cancer but has not been studied to date in FFPE samples. (nih.gov)
Posttranslational modification (PTM), such as tyrosinephosphorylation (pTyr, pY) of proteins, is essential for several cellular functions and pathophysiological processes. (navinci.se)
Enrichment1
A combination of optimized magnetic bead-based sample processing, optimized pTyr enrichment strategies, and tandem mass tag multiplexing enabled in-depth coverage of pTyr signaling networks from small amounts of input material. (nih.gov)
Sites1
Here we describe a method for highly sensitive, quantitative analysis of pTyr signaling networks, with hundreds of sites quantified from one to two 10-μm sections of FFPE tissue specimens. (nih.gov)
Analysis2
In addition, pTyr analysis that may otherwise inform therapeutic interventions for patients has been limited by the requirement for large amounts of frozen tissue. (nih.gov)
pTyr analysis of FFPE tissue sections from breast cancer tumors as well as lung cancer tumors highlighted patient-specific oncogenic driving kinases, indicating potential targeted therapies for each patient. (nih.gov)
Cell3
Macrophage CSF-1R pTyr-721 signaling promotes cell motility and enhancement of tumor cell invasion in vitro. (elsevierpure.com)