Abstract
INTRODUCTIONIn this protocol, radiolabeled proteins are separated using SDS-PAGE and digested in-gel. The phosphopeptides are subjected to two-dimensional LC and identified by MALDI-MS and MALDI-PSD. The identification of eight tyrosine phosphorylation sites in the human Gab-1 protein is provided as an example of the method's utility. The expression and phosphorylation of human Gab-1 with the insulin receptor kinase in vitro are summarized as an example of this approach to identifying phosphorylated tyrosine residues.
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