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Effect of ligand‐affinity differences of human hemoglobin variants on electrophoretic behavior and their isolation and functional characterization
Author(s) -
Rochette Jacques,
Deburgrave Nathalie,
Bohn Brigitte,
Dodé Catherine,
Poyart Claude,
Krishnamoorthy Rajagopal
Publication year - 1989
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.1150101210
Subject(s) - hemoglobin , hemoglobin variants , electrophoresis , isoelectric focusing , chemistry , bohr effect , ligand (biochemistry) , biochemistry , heme , mutant , chromatography , biophysics , biology , receptor , gene , enzyme , oxygen–haemoglobin dissociation curve
A natural sulfated polysaccharide (agaropectin), contained in crude agar, can be used as a medium for electrophoretic separation of hemoglobin mutants, constituting a particular class of protein‐ligand interactions. Mutations which either modify the electrostatic charge at the surface of the hemoglobin molecule or not, have been studied according to their putative interaction with the medium. Using conformational specificities of the hemoglobin molecule, we have also demonstrated that isoelectric focusing on a polyacrylamide gel in the absence of heme ligands represents a useful, convenient and rapid procedure for isolating silent Hb variants in their native form, provided that they exibit an abnormal Bohr effect.