Phosphorylation can reveal essential cell functions, such as cell differentiation, signal transduction, metabolic maintenance and cell division. The aim of this study was to investigate phosphorylated protein expression changes during neuronal lineage differentiation from hESCs. To measure the phosphorylated protein expression change during neuronal differentiation, we performed a comparative phosphoproteome analysis using 2-DE after MALDI-TOF MS and an MS/MS protein identification method, making a comparison between neural lineage differentiating cells and normal embryoid bodies (EBs) differentiated from human embryonic stem cells (hESCs) and profiling constituent phosphorylated proteins. Of 36 differentially expressed protein spots, 12 spots were shown to be up-regulated in differentiating neural cells. Specifically, the 7 up-regulated proteins of the 12 have potential roles in neuronal differentiation or neuronal damage recovery, including ACTB, heterogeneous nuclear ribonucleoprotein A2B1 (hnRNP A2B1), heterogeneous nuclear ribonucleoprotein L (hnRNP L), SET, chaperonin-containing TCP-1, vimentin and voltage-dependent anion channel protein 1 (VDAC1). These proteins are discussed further below.
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