title: Intersubunit circular permutation of human hemoglobin.
abstract: For many years, human hemoglobin (Hb) isolated from erythrocytes has been investigated as a potential oxygen delivery therapeutic. Advantages with respect to the need for blood typing were balanced with various undesirable properties of cell-free Hb, including cost, overall oxygen affinity, alterations in cooperativity, and ready dissociation into toxic dimeric species. The use of total gene synthesis has resulted in very high levels of functional human Hb expression in Escherichia coli, but there remains a desire for effecting the crosslinking of the hemoglobin tetramer and providing for ready means for increasing the globular molecular weight. In this communication, we report a novel method for linking alpha chains. By circularly permuting one alpha sequence, the second alpha chain in the Hb tetramer can be linked with glycine residues to form 2 bridges across the central cavity. The second alpha chain thus presents its amino and carboxyl termini on a solvent exposed surface, providing for additional polymerization of oxygen-carrying subunits or attachment of any other peptide-based therapeutic.
authors.affiliation: Beckman Institute for Advanced Science and Technology and the Department of Biochemistry, University of Illinois, Urbana, IL 61801, USA.
authors.forename: Kevin E
authors.initials: KE
authors.lastname: Sanders
authors.forename: John
authors.initials: J
authors.lastname: Lo
authors.forename: Stephen G
authors.initials: SG
authors.lastname: Sligar
mesh_terms: D001802:Blood Substitutes / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D015195:Drug Design; D004355:Drug Stability; D005822:Genetic Vectors; D005914:Globins / Q000737:chemistry / Q000235:genetics; D006454:Hemoglobins / Q000138:chemical synthesis* / Q000235:genetics; D006801:Humans; D016296:Mutagenesis; D010100:Oxygen / Q000378:metabolism; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis / Q000235:genetics
chemical_list: D001802:Blood Substitutes; D006454:Hemoglobins; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; C079129:hemoglobin polymer; D005914:Globins; D010100:Oxygen