Comparative studies of recombinant human granulocyte-colony stimulating factor, its Ser-17 and (His)6-tagged forms interaction with metal ions by means of immobilized metal ion affinity partitioning. Effect of chelated nickel and mercuric ions on extraction and refolding of proteins from inclusion bodies.

摘要: Abstract The chelation capability of the reactive dye Light Resistant Yellow 2KT towards metal ions, particularly mercury(II) was evaluated in pH range 5.0–7.0, and it shown that dye–Hg(II) complex has a free site for interaction with human recombinant granulocyte-colony stimulating factor (rhG-CSF) from Escherichia coli . Affinity partitioning three rhG-CSF forms – native, rhG-CSF[Cys 17 →Ser ] (His) 6 –rhG-CSF studied aqueous two-phase systems, which contained ions Cu(II), Ni(II) Hg(II) chelated by dye–poly(ethylene glycol) at 5.0 7.0, presence or absence many selected agents. It determined, exhibited stronger hexahistidine-tagged protein form, while extraction power Cu(II) found to be comparable order magnitude all 7.0. A comparative study both its 7.0 revealed possible direct between unpaired Cys-17 rhG-CSF. inclusion body extract thus explaining efficiency targeted proteins renaturation gained upon their interactions ions.