VI 16832

VI 16832 is a broad spectrum Type I kinase inhibitor which can be used as an enrichment tool for the comparative expression analysis of protein kinases in different cancer cell lines.

Signaling Pathways >> Others >> Others

Research Areas >> Cancer

Type I kinase[1]

VI 16832 is a broad spectrum Type I kinase inhibitor[1]. Phosphoproteomics analysis of VI 16832-enriched fractions from MV4-11, HCT116, or 435S cells results in more than 8500 phosphopeptide identifications. These translate into almost 1700 distinct phosphopeptide species derived from 212 different members of the protein kinase superfamily. Analysis of VI 16832-retained proteins from the three cancer cell lines considerably increases the overall number of identified phosphorylation sites on protein kinases. Considering the sum of all phosphopeptide intensities as a measure for VI 16832-enriched protein amount, more than 80% is derived from protein kinases[2].

Cells are lysed in a volume of 35 to 40 mL per experiment. The protein amounts of the starting extracts used in the first and second experiments are: 435S, 85 and 120 mg; HCT116, 240 and 175 mg; MV4-11, 180 and 120 mg. Lysates are adjusted to 1 M NaCl prior to loading onto the VI 16832 column at a flow rate of 0.07 mL/min. Subsequent washing and elution steps are performed. Protein-containing elution fractions are lyophilized, resuspended in one tenth of the initial volume, and then desalted by protein precipitation prior to gel electrophoresis[2].

[1]. Kyla. A. L. Collins, et al. Proteomic Analysis Defines Kinase Taxonomies Specific for Subtypes of Breast Cancer. Apr. 1, 2017.

[2]. Felix S. Oppermann, et al. Large-scale Proteomics Analysis of the Human Kinome. Mol Cell Proteomics. 2009 Jul; 8(7): 1751-1764.

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