Chiral copper-chelate complexes alter selectivities in metal affinity protein partitioning.
Proteins can be distinguished by exploiting complementarity between a histidine's microenvironment and a metal-chelate ligand in metal-affinity separations. The partitioning behavior of three myoglobins was investigated in aqueous two-phase polyethylene glycol-dextran systems containing polyethylene glycol derivatized with Cu(II) complexes of the L- and D-isomers of methionine and aspartate. TSK chromatographic supports derivatized with the methionine complexes were used to study retention of these proteins in metal-affinity chromatography. In the partitioning studies, the amino acid metal chelates exhibit selectivities for the myoglobins that are different from that of Cu(II)-iminodiacetate. Significant differences in selectivity based on the chiral nature of the amino acid complexes were also observed. The chromatographic selectivities of the chelating ligands exhibit little variation, however, suggesting that interactions occurring in solution but not on a surface play an important role in protein binding to the Cu(II)-amino acid-PEG complexes. In solution, the Cu(II)-amino acid complexes are sensitive probes of the microenvironments of surface histidines. The choice of the metal chelate affinity ligand offers a powerful means by which the selectivity of metal-affinity separations can be altered.
Wuenschell
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GE
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena 91125.
Wen
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Todd
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Shnek
D
D
Arnold
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FH
D000818:Animals; D002614:Chelating Agents / Q000737:chemistry; D002846:Chromatography, Affinity; D003300:Copper / Q000737:chemistry; D006736:Horses; D008670:Metals / Q000737:chemistry*; D008970:Molecular Weight; D009211:Myoglobin / Q000032:analysis; D011092:Polyethylene Glycols; D011506:Proteins / Q000737:chemistry*; D012116:Resins, Plant; D012756:Sheep; D014907:Whales
D002614:Chelating Agents; D008670:Metals; D009211:Myoglobin; D011506:Proteins; D012116:Resins, Plant; D011092:Polyethylene Glycols; D003300:Copper