Microcalorimetric study of the effect of hexa-histidine tag and denaturant on the interaction mechanism between protein and metal-chelating gel

Fu Yung Lin, Wen Yih Chen, Hsiu Mei Chen

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

A recombinant protein, Schistosoma japonicum glutathione-S-transferase (SjGST), was fused with a C-terminal hexa-histidine tag to obtain SjGST/His. Both proteins were used to probe the interaction mechanisms with the metal ions immobilized on chromatography gels. Isothermal titration calorimetry was used to directly measure the adsorption enthalpies (Δ Hads) of both proteins with Ni-NTA and TALON (Co2+) commercial affinity resins, under the conditions of with and without the presence of a denaturant. The result reveals that SjGST/His had a lower Δ Hads value with Ni-NTA than did SjGST, mainly attributed to the formation of more coordination bonds with or a stronger binding with Ni-NTA. Furthermore, the difference between the Δ Hads values of SjGST/His onto TALON under the nature and denaturing conditions were insignificant, implying that the binding topography of the hexa-histidine tail with immobilized Co2+ was not significantly changed with the presence of a denaturant. In addition, this study shows that the proposed binding models and the directly measured adsorption heat can be combined to elucidate the difference in the interaction mechanisms of SjGST/His adsorption onto those two adsorbents from a thermodynamic perspective.

Original languageEnglish
Pages (from-to)333-339
Number of pages7
JournalJournal of Colloid and Interface Science
Volume238
Issue number2
DOIs
StatePublished - 15 Jun 2001

Keywords

  • Adsorption enthalpy
  • Immobilized metal ion affinity chromatography, IMAC
  • Isothermal titration calorimetry, ITC
  • Poly-histidine tag
  • Protein adsorption
  • Schistosoma japonicum glutathione-S-transferase

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