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Kinase Inhibitor Profiling of TrkA- and TrkB- Expressing Neuroblastoma Cell Lines Using Desthiobiotin Nucleotide Probes

Reputable Mentor II
Reputable Mentor II
Bomgarden RD, Etienne CL, Viner R, Rosenblatt MM, Cichon E, Rogers JC.
Application Note 515
Here, we examined the relative expression and inhibition of kinases in SH-SY5Y neuroblastoma cells stably expressing TrkA or TrkB. We employed a proteomic approach using desthiobiotin nucleotide probes to specifically capture and profile the kinome of each cell line using mass spectrometry to identify labeled kinase active-site peptides (Figure 1). In addition, we assessed staurosporine and lestaurtinib inhibition of protein kinases using kinase active-site probes in combination with Tandem Mass Tag (TMT) reagents and validated our results using a parallel Western blot workflow (Figure 2). Finally, because this approach allows for the specifi c enrichment of adenine nucleotide binding proteins of interest, an SRM-based workflow could be used to enable targeted quantification as an alternative means of validating the relative expression and inhibition of the kinases, and may be employed as a follow-up to this study.

Thermo Fisher Scientific
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Last update:
‎10-15-2021 11:00 AM
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