Effect of covalent labeling of dextran-benzenehexacarboxylate on hemoglobin.
The covalent fixation of benzenehexacarboxylate (BHC) onto dextran was carried out according to several reaction schemes. The polyanionic polymers thus synthesized were capable of decreasing the oxygen affinity of hemoglobin by specifically interacting with the 2,3-diphosphoglycerate (2,3-DPG) binding site of the protein. The intensity of this effect was correlated to both the chemical structure of the polyanionic polymers and the BHC content in polymer. The polyanionic polymer, containing 0.035 mol BHC/g and presenting no cross-linking between its polymer chains, possessed the best effector properties. These properties were used to direct the covalent fixation of the dextran-benzenehexacarboxylate onto the phosphate binding site of the protein. The resulting hemoglobin was mainly substituted at the same time by one or more linked BHC onto both alpha beta dimers in the vicinity of the 2,3-DPG site. Thus, the modification of hemoglobin led to an increase in the hydrodynamic volume of each dimer sufficient to limit the diffusion of the conjugates through the kidney membrane, even if the conjugates had dissociated into alpha beta dimers. Compared to that of free hemoglobin, the oxygen affinity of the conjugates was significantly decreased. This type of covalent conjugate exhibited general properties quite suitable for use as blood substitutes.
Sacco
D
D
Laboratory of Macromolecular Physical Chemistry, URA-CNRS-494, ENSIC, Nancy, France.
Dellacherie
E
E
Prouchayret
F
F
D019794:2,3-Diphosphoglycerate; D001565:Benzoates / Q000138:chemical synthesis; D001665:Binding Sites; D003911:Dextrans / Q000138:chemical synthesis; D004163:Diphosphoglyceric Acids; D006454:Hemoglobins / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010108:Oxyhemoglobins / Q000737:chemistry* / Q000378:metabolism
D001565:Benzoates; D003911:Dextrans; D004163:Diphosphoglyceric Acids; D006454:Hemoglobins; D010108:Oxyhemoglobins; D019794:2,3-Diphosphoglycerate; C087947:dextran benzenehexacarboxylate