南京林业大学学报(自然科学版) ›› 2015, Vol. 39 ›› Issue (01): 149-154.doi: 10.3969/j.issn.1000-2006.2015.01.027

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蛋白质组学研究中磷酸化蛋白质(肽)富集策略及展望

甄 艳,李春映,陆 叶,施季森*   

  1. 南京林业大学,林木遗传与生物技术省部共建教育部重点实验室,江苏 南京 210037
  • 出版日期:2015-01-31 发布日期:2015-01-31
  • 基金资助:
    收稿日期:2013-10-17 修回日期:2014-06-26
    基金项目:江苏省高校自然科学重大研究项目(12KJA220002); 博士点基金优先发展领域项目(20113204130002); 江苏高校优势学科建设工程资助项目(PAPD)
    第一作者:甄艳, 副教授,博士。*通信作者:施季森,教授。E-mail:jshi@njfu.edu.cn。
    引文格式:甄艳,李春映,陆叶,等. 蛋白质组学研究中磷酸化蛋白质(肽)富集策略及展望[J]. 南京林业大学学报:自然科学版,20

Enrichment strategies and prospects of phosphoprotein(phosphopetide)in phosphoproteomics

ZHEN Yan, LI Chunying, LU Ye, SHI Jisen*   

  1. Key Laboratory of Forest Genetics &
    Biotechnology of Ministry of Education, Nanjing Forestry University, Nanjing 210037, China
  • Online:2015-01-31 Published:2015-01-31

摘要: 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。

Abstract: 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.

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