title: Effect of ligand-affinity differences of human hemoglobin variants on electrophoretic behavior and their isolation and functional characterization.
abstract: 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 exhibit an abnormal Bohr effect.
authors.affiliation: Biochimie Génétique, CHU Cochin, Paris, France.
authors.forename: J
authors.initials: J
authors.lastname: Rochette
authors.forename: N
authors.initials: N
authors.lastname: Deburgrave
authors.forename: B
authors.initials: B
authors.lastname: Bohn
authors.forename: C
authors.initials: C
authors.lastname: Dodé
authors.forename: C
authors.initials: C
authors.lastname: Poyart
authors.forename: R
authors.initials: R
authors.lastname: Krishnamoorthy
mesh_terms: D002118:Calcium; D004563:Electrochemistry; D004586:Electrophoresis / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D014644:Genetic Variation; D006418:Heme; D006455:Hemoglobins, Abnormal / Q000302:isolation & purification*; D006801:Humans; D007525:Isoelectric Focusing; D008024:Ligands; D008274:Magnesium
chemical_list: D006455:Hemoglobins, Abnormal; D008024:Ligands; D006418:Heme; D008274:Magnesium; D002118:Calcium