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Immobilized metal ion affinity gel electrophoresis: Quantification of protein affinity to transition metal chelates
Author(s) -
Baek WonOk,
Haupt Karsten,
Colin Carole,
Vijayalakshmi Mookambeswaran A.
Publication year - 1996
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150170310
Subject(s) - affinity electrophoresis , chelation , iminodiacetic acid , chemistry , affinity chromatography , agarose , metal , metal ions in aqueous solution , chromatography , ligand (biochemistry) , affinities , electrophoresis , gel electrophoresis , inorganic chemistry , organic chemistry , stereochemistry , biochemistry , enzyme , receptor
This paper describes some recent advances in the methodology of immobilized metal ion affinity gel electrophoresis. Four different ways to incorporate metal chelate ligands in agarose and polyacrylamide‐based electrophoresis gels are evaluated, a new polymerizable metal chelating ligand, allyl–2‐hydroxy‐3‐( N,N ‐dicarboxymethyl)amino‐propyl ether, is introduced, and the determination of affinity constants described. The affinities of model proteins (ribonucleases A and B and cytochromes c from different species) for the transition metal chelate iminodiacetic acid‐Cu(II) were studied. The results were found to be in agreement with literature data on immobilized metal ion affinity chromatography, and the polymer nature and the different chemistries used influenced the affinity only quantitatively, keeping the basic mechanisms of interaction unchanged.