Streamlined Membrane Proteome Preparation for Shotgun Proteomics Analysis with Triton X-100 Cloud Point Extraction and Nanodiamond Solid Phase Extraction
作者: Minh PhamTing-Chun WenHung-Cheng LiPei-Hsuan HsiehYet-Ran ChenHuan-Cheng ChangChau-Chung Han
作者单位: 1Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei 10617, Taiwan
2Institute of Biotechnology, Vietnam Academy of Science & Technology, Hanoi 10600, Vietnam
3Agricultural Biotechnology Research Center, Academia Sinica, Taipei 11529, Taiwan
刊名: Materials, 2016, Vol.9 (5)
来源数据库: Multidisciplinary Digital Publishing Institute
DOI: 10.3390/ma9050385
关键词: nanodiamonddiamond-enhanced proteolytic digestionmembrane proteomicsmass spectrometrysalt-induced phase separationcloud point extraction
原始语种摘要: While mass spectrometry (MS) plays a key role in proteomics research, characterization of membrane proteins (MP) by MS has been a challenging task because of the presence of a host of interfering chemicals in the hydrophobic protein extraction process, and the low protease digestion efficiency. We report a sample preparation protocol, two-phase separation with Triton X-100, induced by NaCl, with coomassie blue added for visualizing the detergent-rich phase, which streamlines MP preparation for SDS-PAGE analysis of intact MP and shot-gun proteomic analyses. MP solubilized in the detergent-rich milieu were then sequentially extracted and fractionated by surface-oxidized nanodiamond (ND) at three pHs. The high MP affinity of ND enabled extensive washes for removal of salts, detergents,...
全文获取路径: MDPI 

  • stream 
  • digestion 消化
  • removal 消去
  • detergent 清洁剂
  • achieved 获得的
  • hydrophobic 疏水的
  • higher 
  • ensure 担保
  • efficiency 效率
  • weight