The contribution of Cys17 and surface-exposed histidine residues in rhG-CSF interaction with Cu(II), Ni(II) and Hg(II) ions chelated by Light Resistant Yellow 2KT-polyethylene glycol derivative was evaluated in aqueous two-phase systems composed of polyethylene glycol (PEG) and dextran. It was determined that His43, His52, His156 and His170 residues are involved in protein interaction with chelated Cu(II) ions. Protein interaction with chelated Ni(II) is governed by His52 and His170 residues, though Cys17 is also involved. The contribution of Cys17 side chain is dominant in the interaction between rhG-CSF and chelated Hg(II) ions. The direct interaction between chelated Hg(II) ions and the –SH group of protein was determined for the first time. Based on the study of the interaction between rhG-CSF and chelated metal ions, rhG-CSF was successfully refolded from inclusion bodies in aqueous two-phase systems PEG-dextran containing chelated Ni(II) or Hg(II) ions for the first time. The refolding of rhG-CSF (C17S) in these systems was more effective compared to that of intact rhG-CSF. The dependence of refolding efficiency of rhG-CSF (C17S) in two-phase systems containing chelated metal ions on the number of histidine mutations was evaluated. It was determined that the refolding efficiency of protein in the systems containing chelated Ni(II) is inversely proportional to the number of histidine mutations. The affinity of purified rhG-CSF (C17S) and its histidine mutants for... [to full text]
Identifer | oai:union.ndltd.org:LABT_ETD/oai:elaba.lt:LT-eLABa-0001:E.02~2005~D_20051116_154617-68277 |
Date | 16 November 2005 |
Creators | Zaveckas, Mindaugas |
Contributors | Maruška, Audrius, Sereikaitė, Jolanta, Žvirblienė, Aurelija, Matulis, Daumantas, Meškys, Rolandas, Pesliakas, Henrikas, Kulys, Juozas, Lagunavičius, Arūnas, Vidžiūnaitė, Regina, Vilnius Gediminas Technical University |
Publisher | Lithuanian Academic Libraries Network (LABT), Vilnius Gediminas Technical University |
Source Sets | Lithuanian ETD submission system |
Language | English |
Detected Language | English |
Type | Doctoral thesis |
Format | application/pdf |
Source | http://vddb.library.lt/obj/LT-eLABa-0001:E.02~2005~D_20051116_154617-68277 |
Rights | Unrestricted |
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