[Recombinant human hemoglobin with low oxygen affinity: additional effects of two mutations].
The search for human Hb variants exhibiting a low oxygen affinity without requiring 2,3-diphosphoglycerate, together with a low oxygation rate, is of an increased interest in the view of producing an artificial oxygen carrier. We have synthesized the recombinant Hb beta 41Phe-->Tyr (rHb beta F41Y) which exhibits a low oxygen affinity due to the stabilization of the deoxy state of tetrameric Hb [1]. Interestingly, the autooxydation rate for this mutant is similar to that for Hb A. We have associated the mutation beta F41Y with the naturally occurring beta 82Lys-->Asp substitution (Hb Providence) known to be responsible for a low oxygen affinity [2]. The second-site mutation further decreases the oxygen affinity of the rHb beta F41Y. The effects of the beta F41Y and K82D mutations are additive, resulting in a four fold decrease in oxygen affinity of the artificial mutant Hb beta F41Y-K82D, compared to Hb A. In spite of the marked decrease in oxygen affinity, the autooxydation rate is 2- to 3 fold larger than that of Hb A. These data show that it is possible to adjust the oxygen binding properties of human Hb by using protein engineering methods. Because of the low oxygen affinity coexisting with a moderately increased autooxydation rate, this variant is a good candidate for the development of a Hb-based oxygen carrier.
Baudin
V
V
INSERM U229, Hôpital de Bicêtre, Le Kremlin Bicêtre.
Dumoulin
A
A
Poyart
C
C
Pagnier
J
J
D000494:Allosteric Regulation; D001224:Aspartic Acid / Q000737:chemistry; D006454:Hemoglobins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008239:Lysine / Q000737:chemistry; D009154:Mutation; D010084:Oxidation-Reduction; D010100:Oxygen / Q000097:blood*; D010649:Phenylalanine / Q000737:chemistry; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D014443:Tyrosine / Q000737:chemistry
D006454:Hemoglobins; D011994:Recombinant Proteins; D001224:Aspartic Acid; D014443:Tyrosine; D010649:Phenylalanine; D008239:Lysine; D010100:Oxygen
Hémoglobine humaine recombinante de faible affinité pour l'oxygène: effets additionnels de deux mutations.