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For example, the mapping of phosphorylation sites and analyses of phosphorylation site mutants have tremendously contributed to our knowledge of different cellular processes.
Next, we performed a similar mapping of phosphorylation sites on human hASF1a and hASF1b with baculovirus expressed hTLK2 (Figure 1A).
The Atf1-11M-HA proteis is not phosphorylated by Spc1 or by any other protein kinase, as judged by two-dimensional mapping of phosphorylation events [40].
One problem in large-scale mapping of phosphorylation sites is that this method alone cannot reveal direct kinase-substrate relationships.
In recent years, advances in mass spectrometry (MS -based protein analysis technologies, coMS -basedth proteinpeptide enrichment methods, panalysis way for large-scale mapping of phosphorylatechnologiesn vivo [ 13, 18, 23].
However, since the class I DYRKs (DYRK1A and DYRK1B) can also act as priming kinases for GSK3β, attribution of DYRK1A as a T286 kinase requires careful mapping of phosphorylation in vitro and in vivo.
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Large-scale mapping of phosphorylations that occur in response to diverse environmental signals has become an indispensable method for unraveling plant regulatory networks [ 17- 22].
Furthermore, a new strategy called CEASAR, based on functional protein microarrays and bioinformatics, has been developed to construct a high-resolution map of phosphorylation networks that connects 230 kinases to 2591 phosphorylation sites in 652 substrates (28).
Analysis of cell-cycle averaged phosphorylation of APC subunits resulted in mapping of several phosphorylation sites (Fig. 4C).
The increased stability of the modified peptides allowed mapping of the phosphorylation site by tandem mass spectrometry.
The peptides were subjected to HILIC fractionation prior to MS analysis, which allows for in-depth mapping of protein phosphorylation sites in a complex sample.
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