sh2 domain  (Proteintech)

Journal: bioRxiv

Article Title: SH2 scan : Mapping SH2 domain-ligand binding selectivity for inhibitors and degraders

doi: 10.1101/2025.07.09.663991

Figure Lengend Snippet:

Article Snippet: J.A.B. is a co-inventor on a pending patent application filed by Eurofins DiscoverX, LLC for SH2 domain competition binding assays.

Techniques: Construct, Mutagenesis, Phospho-proteomics

Journal: bioRxiv

Article Title: SH2 scan : Mapping SH2 domain-ligand binding selectivity for inhibitors and degraders

doi: 10.1101/2025.07.09.663991

Figure Lengend Snippet: a , An SH2 domain-containing protein construct (blue) fused to the NFκB DNA binding domain (pink) is tagged with an exogenous double-stranded DNA (dsDNA) probe. This construct is incubated with a capture ligand (red) immobilized on magnetic beads (green). b , In the presence of a competitor compound (yellow), less tagged protein is captured on the beads. The protein remaining on the beads is eluted using a high concentration of sodium phenyl phosphate, a generic competitor of phosphopeptide binding to SH2 domains . After elution, a lower qPCR signal is obtained at the end of the assay. In the presence of a non-competitor compound ligand (brown), more tagged protein is captured on the beads and a high qPCR signal is observed. c , Representative primary screening data for the STAT3 construct are shown. Compounds were tested at 10 μM and the hit cutoff for the compound screen was set at ≤35% of the average signal of the DMSO control wells for each construct (dashed line). Percent assay signal for each compound is expressed as the mean of at least two independent technical replicates from at least one independent experiment, ± standard deviation. d , K D data for selected validated hits for the STAT3 construct are shown. Compounds were tested in dose-response, and the data points were fit to the Hill equation (see Methods). Data are presented as mean percent assay signal from at least four independent technical replicates collected over two independent experiment, ± standard deviation.

Article Snippet: J.A.B. is a co-inventor on a pending patent application filed by Eurofins DiscoverX, LLC for SH2 domain competition binding assays.

Techniques: Construct, Binding Assay, Incubation, Magnetic Beads, Concentration Assay, Phospho-proteomics, Control, Standard Deviation

Journal: bioRxiv

Article Title: SH2 scan : Mapping SH2 domain-ligand binding selectivity for inhibitors and degraders

doi: 10.1101/2025.07.09.663991

Figure Lengend Snippet: Percent of DMSO control values were measured for each compound tested in this study and mapped as dots on a phylogenetic tree containing all 120 canonical human SH2 domains. Each dot plotted in a diagram represents a mean value collected from at least two independent technical replicates collected over at least one independent experiment, ± standard deviation. The screening results for the two SH2 domains of SYK and ZAP70 are mapped as equivalently sized dots, given that the SH2 domains for these targets are expressed in tandem constructs, and any competition observed cannot be unequivocally assigned to one or the other SH2 domain.

Article Snippet: J.A.B. is a co-inventor on a pending patent application filed by Eurofins DiscoverX, LLC for SH2 domain competition binding assays.

Techniques: Control, Standard Deviation, Construct