title: Improvement of oxygen-carrying properties of human hemoglobin by chemical modification with a benzene hexacarboxylate-monosubstituted polyoxyethylene.
abstract: Benzene hexacarboxylate-monosubstituted polyoxyethylene on contact with Hb decreases its oxygen affinity, probably because it specifically interacts with the amino groups of the phosphate-binding site. This site specificity was used to direct the covalent coupling of this polymeric reagent with hemoglobin, in the vicinity of this beta cleft in order to obtain conjugates with low oxygen affinity and well-defined molecular weight. Such conjugates could thus be regarded as potential candidates for blood substitutes. Covalent fixation of this polymeric site-labeling reagent onto hemoglobin was carried out with the oxy and the deoxy form in the presence of a water soluble carbodiimide. It turns out that the oxygen-binding properties of the resulting hemoglobin derivatives depend on the reaction conditions, yet in all cases the oxygen affinity of the modified protein was lower than that of native hemoglobin and was no longer affected by organic phosphates. These results indicate that phosphate-binding site amines are probably involved in the covalent coupling, although in some conjugates (especially those prepared with high ratios of reagents) other amino groups participate also in the linking to the polymer. Chromatographic analysis and tryptic peptide mapping of some conjugates evidenced that the beta-terminal valine residue was in fact the preferential binding site of hexacarboxylate-monosubstituted polyoxyethylene.
authors.affiliation: Laboratory of Macromolecular Physical Chemistry, URA CNRS, Nancy, France.
authors.forename: E
authors.initials: E
authors.lastname: Dellacherie
authors.forename: M
authors.initials: M
authors.lastname: Léonard
mesh_terms: D001665:Binding Sites; D006454:Hemoglobins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D010100:Oxygen / Q000378:metabolism*; D010108:Oxyhemoglobins / Q000378:metabolism; D010449:Peptide Mapping; D011092:Polyethylene Glycols / Q000494:pharmacology*
chemical_list: D006454:Hemoglobins; D010108:Oxyhemoglobins; D011092:Polyethylene Glycols; D010100:Oxygen