The analysis of protein function has conventionally been done by studying, in depth, a single protein. Recently several groups have initiated efforts to display proteins in two-dimensional array suitable for performing various types of binding studies or assays. In particular, Dr. Snyder's laboratory has developed a microwell format for protein display that has versatility and a range of applications that compare favorably with other formats. This format has been used to create arrays of essentially all yeast kinases and to study their specificity. In the present Phase I application we propose to extend this work by creating an array of 96 human protein kinase catalytic domains. To do this we will express domains in a baculovirus system suitable for multiplexing production, and demonstrate the utility of the microwell format by testing the activity and specificity of the kinases. We will also further decrease the size and increase the density of microwells in arrays, in preparation for preparation of large-scale protein arrays.
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| Mok, Janine; Im, Hogune; Snyder, Michael (2009) Global identification of protein kinase substrates by protein microarray analysis. Nat Protoc 4:1820-7 |
