title: Three-point cross-linking: potential red cell substitutes from the reaction of trimesoyl tris(methyl phosphate) with hemoglobin.
abstract: The symmetrical trifunctional cross-linking reagent trimesoyl tris(methyl phosphate) (3), reacts selectively with amino groups (beta 1Val and beta 82Lys) in the diphosphoglycerate binding site of human hemoglobin A, producing cross-linked tetrameric species in good yield. A major species is triply linked, alpha alpha beta 1(82) greater than B beta 82, where B symbolizes benzene-1,3,5-tricarbonyl. Both this triply linked species and the doubly linked species, alpha alpha beta 1B beta 82, produced from deoxyhemoglobin have a considerably lower oxygen affinity than does native hemoglobin while maintaining a high degree of cooperativity (n50 = 2.4), making them potentially useful as red cell substitutes, in principle delivering twice as much oxygen as whole blood between pO2 = 100 and = 40 Torr. The yield of products indicates that triply and doubly linked species form in parallel so that there are independent routes to each. It is proposed that differences in routes are due to stereoisomerism about the amide bonds which form from reaction of the reagent with the protein.
authors.affiliation: Lash Miller Laboratories, Department of Chemistry, University of Toronto, Ontario, Canada.
authors.forename: R
authors.initials: R
authors.lastname: Kluger
authors.forename: J
authors.initials: J
authors.lastname: Wodzinska
authors.forename: R T
authors.initials: RT
authors.lastname: Jones
authors.forename: C
authors.initials: C
authors.lastname: Head
authors.forename: T S
authors.initials: TS
authors.lastname: Fujita
authors.forename: D T
authors.initials: DT
authors.lastname: Shih
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D001802:Blood Substitutes; D003432:Cross-Linking Reagents / Q000378:metabolism*; D004163:Diphosphoglyceric Acids / Q000378:metabolism; D017707:Erythrocyte Transfusion; D006441:Hemoglobin A / Q000378:metabolism*; D006801:Humans; D046911:Macromolecular Substances; D009682:Magnetic Resonance Spectroscopy; D008433:Mathematics; D008958:Models, Molecular; D008961:Models, Structural; D008962:Models, Theoretical; D009943:Organophosphorus Compounds / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D013055:Spectrophotometry, Infrared
chemical_list: D001802:Blood Substitutes; D003432:Cross-Linking Reagents; D004163:Diphosphoglyceric Acids; D046911:Macromolecular Substances; D009943:Organophosphorus Compounds; D010446:Peptide Fragments; C076346:trimesoyl tris(methyl phosphate); D006441:Hemoglobin A