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. Author manuscript; available in PMC: 2011 Mar 8.
Published in final edited form as: Ann N Y Acad Sci. 2008 Oct;1141:22–57. doi: 10.1196/annals.1441.022

Figure 2.

Figure 2

The use of analog-sensitive kinases can facilitate substrate identification. The analog-sensitive kinase (as-kinase) is designed to incorporate ATP-γS analogs that have a bulky side group (*ATP-γS). Native kinases can incorporate normal ATP-γS but cannot utilize the *ATP-γS. Substrates phosphorylated by the as-kinase can be identified by alkylation with para-nitrobenzylmesylate (PNBM) to create an epitope that is recognized by antibodies specific for the thiophosphate ester tag. The substrates can be isolated by immunoprecipitation or immunoaffinity purification and analyzed by mass spectrometry. Adapted from Reference 276.