Citation Spotlight: Toolsets to Define PTMs for Any Target Protein

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Comparison of BlastR lysis buffer to non-denaturing lysis buffers and detection of total SUMOylation 2/3, acetylation, ubiquitination, and tyrosyl phosphorylation profiles were detected with their respective antibodies.

Recently, Horita et al. examined the post-translational modification (PTM) profile changes in the EGFR→Ras→c-Fos signaling pathway in response to EGF stimulation. PTMs are dynamic and often reversible modifications that alter protein structure and function. While tyrosine phosphorylation (pY) is well-characterized in the EGFR signaling pathway, other PTMs like acetylation (Ac), SUMOylation (SUMO), and ubiquitination (Ub) have not been thoroughly investigated. A novel toolset termed Signal-Seeker™ kits were utilized to investigate the pY, Ac, SUMO, and Ub status of the EGFR signaling axis. All 10 of the previously identified PTMs of the EGFR→Ras→c-Fos signaling pathway were identified, and a novel modification, c-Fos Ac, was also discovered. Importantly, utilizing the toolset enabled investigation of the PTM status of proteins in various cellular compartments that ranged from low to high abundance. The dynamic and endogenous levels of these PTMs were investigated in a single lysis system, providing insight into potential crosstalk between these four PTMs. Cytoskeleton's pY, Ub, SUMO 2/3, and Ac Signal-Seeker™ PTM detection kits (Cat. # BK160, BK161, BK162, and BK163, respectively) were essential reagents in this study, and provide a novel toolset for simple and effective investigation of established and novel PTMs for any target protein. Link to citation:
Horita H. et. al. A simple toolset to identify endogenous post-translational modifications for a target protein: a snapshot of the EGFR signaling pathway. Biosci. Rep. DOI: 10.1042/BSR20170919

Products used in this citation:

Signal-Seeker™ Phosphotyrosine Enrichment Kit (30 assay) (Cat. # BK160)
Signal-Seeker™ Phosphotyrosine Enrichment Kit (10 assay) (Cat. # BK160-S)  Signal-Seeker™ Ubiquitination Enrichment Kit (30 assay) (Cat. # BK161)
Signal-Seeker™ Ubiquitination Enrichment Kit (10 assay) (Cat. # BK161-S) Signal-Seeker™ SUMOylation 2/3 Enrichment Kit (30 assay) (Cat. # BK162)
Signal-Seeker™ SUMOylation 2/3 Enrichment Kit (30 assay) (Cat. # BK162-S) Signal-Seeker™ Acetyl-Lysine Enrichment Kit (30 assay) (Cat. # BK163)
Signal-Seeker™ Acetyl-Lysine Enrichment Kit (30 assay) (Cat. # BK163-S)

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