title: Thermal stability of hemoglobin crosslinked in the T-state by bis(3,5-dibromosalicyl) fumarate.
abstract: Bis(3,5-dibromosalicyl) fumarate was used to crosslink oxyhemoglobin between Lys 82 beta 1 and Lys 82 beta 2 (Walder, J. A., et al. (1979) Biochemistry 18, 4265) and deoxyhemoglobin between Lys 99 alpha 1 and Lys 99 alpha 2 (Chatterjee R.Y., et al. (1986) J. Biol. Chem. 261, 9929). Thermal denaturations demonstrated that alpha crosslinked hemoglobin (alpha 99XLHb A) has the same stability as the beta crosslinked one (beta 82XLHb A). Both alpha and beta crosslinked methemoglobins have a denaturation temperature in 0.9 M guanidine of 57 degrees C compared to 41 degrees C of Hb A. The second product from the T-state crosslinking reaction was found to be crosslinked between the beta chains by chain separation and amino acid analysis. The possible positions for this crosslink are limited to the bisphosphoglycerate binding site in the three-dimensional structure. Its stability is comparable to that of the alpha 99XLHb A or beta 82XLHb A. These modified hemoglobins are potential blood substitutes.
authors.affiliation: Department of Chemistry, Loyola University of Chicago, Chicago, IL 60626.
authors.forename: T
authors.initials: T
authors.lastname: Yang
authors.forename: K W
authors.initials: KW
authors.lastname: Olsen
mesh_terms: D000596:Amino Acids / Q000032:analysis; D001241:Aspirin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000378:metabolism*; D004355:Drug Stability; D005914:Globins / Q000378:metabolism*; D006441:Hemoglobin A / Q000378:metabolism; D006454:Hemoglobins / Q000378:metabolism*; D006801:Humans; D046911:Macromolecular Substances; D011489:Protein Denaturation; D013816:Thermodynamics
chemical_list: D000596:Amino Acids; D003432:Cross-Linking Reagents; D006454:Hemoglobins; D046911:Macromolecular Substances; C021630:bis(3,5-dibromosalicyl)fumarate; D005914:Globins; D006441:Hemoglobin A; D001241:Aspirin