High-throughput Phosphotyrosine Protein Complexes Screening by Photoaffinity-engineered Protein Scaffold-based Forward-phase Protein Array.
作者: Zheng ZhendongChu BizhuKong QianChen XiongKe MiQin YunqiuLu YiFeng ShunTian Ruijun
刊名: Analytical chemistry, 2019
来源数据库: PubMed Journal
DOI: 10.1021/acs.analchem.9b01845
原始语种摘要: Low abundance phosphotyrosine (pTyr)-mediated signaling protein complexes play critical roles in cancer signaling. The precise and comprehensive profiling of these pTyr-mediated protein complexes remains challenging due to their dynamic nature and weak binding affinity. Taking the advantage of the SH2 domains modified with trifunctional chemical probes and genetic mutations (termed Photo-pTyr-scaffold), we developed a Photo-pTyr-scaffold-based forward-phase protein array for specifically capturing by engineered SH2 domain, photoaffinity crosslinking, and antibody-based measuring weak pTyr-mediated protein complexes from complex biological samples in 96-well microplate format. This platform demonstrated good precision for quantitation (R2 = 0.99) and high sensitivity by which only 5 μg of...
全文获取路径: PubMed  (合作)
影响因子:5.695 (2012)

  • throughput 吞吐量
  • engineered 设计的
  • complexes 联合企业
  • protein 蛋白质
  • based 基于
  • screening 筛分
  • demonstrated 探明的储量
  • scaffold 脚手架
  • probes 探针
  • dynamic 动力学的