Protein selectivity with immobilized metal ion-tacn sorbents: chromatographic studies with human serum proteins and several other globular proteins.
The chromatographic selectivity of the immobilized chelate system, 1,4,7-triazocyclononane (tacn), complexed with the borderline metal ions Cu2+, Cr3+, Mn2+, Co2+, Zn2+, and Ni2+ has been investigated with hen egg white lysozyme, horse heart cytochrome c, and horse skeletal muscle myoglobin, as well as proteins present in partially fractionated preparations of human plasma. The effects of ionic strength and pH of the loading and elution buffers on protein selectivities of these new immobilized metal ion affinity chromatographic (IMAC) systems have been examined. The results confirm that immobilized Mn;pl-tacn sorbents exhibit a novel type of IMAC behavior with proteins. In particular, the chromatographic properties of these immobilized M(n+)-tacn ligand systems were significantly different compared to the IMAC behavior observed with other types of immobilized tri- and tetradentate chelating ligands, such as iminodiacetic acid, O-phosphoserine, or nitrilotriacetic acid, when complexed with borderline metal ions. The experimental results have consequently been evaluated in terms of the additional contributions to the interactive processes mediated by effects other than solely the conventional lone pair Lewis soft acid-Lewis soft base coordination interactions, typically found for the IMAC of proteins with borderline and soft metal ions, such as Cu2+ or Ni2+.
Jiang
W
W
Department of Biochemistry and Molecular Biology, Monash University, Victoria, Australia.
Graham
B
B
Spiccia
L
L
Hearn
M T
MT
D000818:Animals; D001798:Blood Proteins / Q000032:analysis / Q000737:chemistry*; D002021:Buffers; D002614:Chelating Agents; D002645:Chickens; D002846:Chromatography, Affinity / Q000379:methods*; D003300:Copper / Q000737:chemistry; D003574:Cytochrome c Group / Q000032:analysis / Q000737:chemistry; D004529:Egg White; D006321:Heart; D006571:Heterocyclic Compounds / Q000737:chemistry; D006736:Horses; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008345:Manganese / Q000737:chemistry; D008670:Metals / Q000737:chemistry; D009113:Muramidase / Q000032:analysis / Q000737:chemistry; D018482:Muscle, Skeletal / Q000737:chemistry; D009211:Myoglobin / Q000032:analysis / Q000737:chemistry; D009994:Osmolar Concentration; D012685:Sepharose / Q000031:analogs & derivatives; D015032:Zinc / Q000737:chemistry
D001798:Blood Proteins; D002021:Buffers; D002614:Chelating Agents; D003574:Cytochrome c Group; D006571:Heterocyclic Compounds; D008670:Metals; D009211:Myoglobin; D008345:Manganese; C075712:1,4,7-triazacyclononane; D003300:Copper; D012685:Sepharose; D009113:Muramidase; D015032:Zinc