Enrichment separation of recombinant human CCR3 using detergent/polymer two-phase system.
Studies on the structure and functions of membrane proteins are impeded by their lability, hydrophobicity, difficulty in purification and low yields. Human chemokine receptor 3 (CCR3) is a G protein-coupled receptor related to allergic diseases. A Triton X-100/PEG20000 two-phase system was employed for enrichment separation of CCR3 over-expressed in E. coli. Optimal CCR3 partitioning with partition coefficient around 8 was obtained at pH 7.0, ionic strength of 0.3 mol/kg and 3 h equilibration time. Total recovery of CCR3 reached 102 +/- 15%, which was much higher than 32 +/- 5% of the normally used ultracentrifugation method. The recovered CCR3 was finally purified by two chromatography steps giving a final protein of 87 kDa.
Ge
Baosheng
B
Center for Bioengineering and Biotechnology, China University of Petroleum, 266555, Qingdao, Shandong, People's Republic of China. gebaosheng@upc.edu.cn
Li
Wei
W
Yin
Yu
Y
D001709:Biotechnology / Q000379:methods*; D003902:Detergents / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D017830:Octoxynol / Q000494:pharmacology*; D009994:Osmolar Concentration; D011092:Polyethylene Glycols / Q000494:pharmacology*; D054397:Receptors, CCR3 / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification*; D013997:Time Factors; D014461:Ultracentrifugation
C515350:CCR3 protein, human; D003902:Detergents; D054397:Receptors, CCR3; D011994:Recombinant Proteins; D011092:Polyethylene Glycols; D017830:Octoxynol