Sulfhemoglobin. Properties of partially sulfurated tetramers.
The properties of sulfhemoglobin (sulfHb) were investigated using disc gel isoelectric focusing and optical spectrophotometry. Laboratory-prepared samples, which contained a high yield of sulfHb (70-85%), and a patient-derived sample, which contained a low yield (12%), contain a tetrameric population that reflects a random distribution of modified (sulfurated) subunits. Hybrid tetramers, i.e. those containing sulfurated and unmodified subunits, were resolved from fully sulfurated and unmodified hemoglobin upon electrofocusing of ferric or ferrous sulHb samples. The electrophoretic differences between ferric sulfHb and metHb arise from a difference in the pK for the met to hydroxymet conversion of sulfurated subunits (Carrico, R., Peisach, J., and Alben, J. O. (1978) J. Biol. Chem. 253, 2386-2391). Both partially and fully sulfurated tetramers, when in the deoxy form and when in the fully ligated CO form, co-focus with their unmodified deoxy-Hb and Hb CO counterparts. Thus, these sulfurated tetramers exhibit normal ligand-dependent ionization of Bohr protons. In air-equilibrated samples, hybrid tetramers are partially ligated as a result of the reduced O2 affinity of the sulfurated subunits. These partially ligated tetramers exhibit a pI intermediate between oxy-Hb and deoxy-Hb due to the fractional ionization of Bohr protons. The partially sulfurated, partially ligated tetramers as well as deoxy-sulfHb tetramers were found to bind 2,3-diphosphoglycerate in disc gels. Despite the preservation of these heterotropic effects, which are confirmed in CO binding studies in solution, fully sulfurated tetramers recovered using preparative isoelectric focusing exhibit little or no cooperativity.
D002248:Carbon Monoxide / Q000097:blood; D006801:Humans; D007525:Isoelectric Focusing; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D011485:Protein Binding; D013053:Spectrophotometry; D013435:Sulfhemoglobin / Q000302:isolation & purification / Q000378:metabolism*
D046911:Macromolecular Substances; D002248:Carbon Monoxide; D013435:Sulfhemoglobin