title: Covalent fixation of polymer-linked benzene hexacarboxylate onto human haemoglobin.
abstract: The reaction of human deoxy and oxyhaemoglobin with a macromolecular effector, monomethoxypolyoxyethylene-linked benzene hexacarboxylate, in the presence of a water soluble carbodiimide, produces under defined conditions, the same conjugates preferentially acylated at the two valines beta 1. The oxygen affinity of both these conjugates is decreased by approximately 5-fold compared with that of native Hb (at pH 7.2, in 0.05 M Tris buffer, 25 degrees C, P50: 20.1 and 20.7 Torr versus about 4 Torr for Hb). This difference appears to be due to an overstabilization of the T state probably together with a decrease of the oxygen affinity of the R state. Addition of IHP to the conjugate solutions does not influence the P50 but addition of IHP to the reaction mixtures before the coupling limits the substitution of Hb by the macromolecular effector, to 20% (instead of 100% in absence of IHP). The cooperativity curve is shifted to the right with an Nmax of 3 at about 90% oxygen saturation, which corresponds to a potential release of 48% of oxygen at pH 7.2, 25 degrees C, between 100 and 40 Torr, compared with 40% for blood. Such kinds of conjugates especially those obtained from oxyhaemoglobin which are easily prepared, could be of a great interest as non-diffusing oxygen carriers in transfusional and perfusional fluids.
authors.affiliation: Laboratory of Macromolecular Physical Chemistry, URA CNRS 494, ENSIC, Nancy, France.
authors.forename: M
authors.initials: M
authors.lastname: Léonard
authors.forename: E
authors.initials: E
authors.lastname: Dellacherie
mesh_terms: D001565:Benzoates / Q000378:metabolism*; D001665:Binding Sites; D006454:Hemoglobins / Q000378:metabolism*; D006801:Humans; D010100:Oxygen / Q000378:metabolism; D010449:Peptide Mapping; D011092:Polyethylene Glycols; D011108:Polymers; D011485:Protein Binding
chemical_list: D001565:Benzoates; D006454:Hemoglobins; D011108:Polymers; D011092:Polyethylene Glycols; C028210:monomethoxypolyethylene glycol; C015339:mellitic acid; D010100:Oxygen