蛋白质组学研究中磷酸化蛋白质(肽)富集策略及展望

Enrichment strategies and prospects of phosphoprotein(phosphopetide)in phosphoproteomics

蛋白质磷酸化是细胞信号转导途径中重要的调控过程,但磷酸化蛋白质(肽)丰度低、化学计量低、易降解等限制了磷酸化蛋白质(肽)的研究。磷酸化蛋白质(肽)的制备是蛋白质磷酸化研究成功的关键步骤。笔者综述了磷酸化蛋白质(肽)的一些富集策略:基于抗体技术、亲和性标签磷酸化蛋白质富集策略,基于固相金属亲和层析、金属氧化亲和层析、IMAC连续洗脱、离子交换层析、亲水相互作用层析的磷酸化肽富集策略,熟练掌握及综合应用这些磷酸化蛋白质(肽)的富集策略,对于研究细胞信号转导途径中的调控因子具有重要作用。

Protein phosphorylation is an important regulation way in cellular signaling pathways. However, it was difficult to study on phosphoprotein/phosphopetide because of the low abundances, low stoichiometry, and easily to degrade in biological environment. The ideal sample preparation was one of the key steps for protein phosphorylation research. The strategy of phosphoprotein/phosphopetide enrichment with antibody, biotin-tagging approach, phosphopetide enrichment with immobilized metal affinity chromatography, metal oxide affinity chromatography, sequential elution from IMAC, ion exchange chromatography, and the hydrophilic interaction liquid chromatography were discussed in this review, respectively. The importance and application of the phosphorylated proteins/peptides enrichment strategy had been proposed for the further study on regulating factors during cellular signaling transduction pathways.