Hemin binding to serum proteins and the catalysis of interprotein transfer.
The reaction of hemin (Hm) with human hemopexin (Hx) has been studied in a mixed dimethyl sulfoxide (Me2SO)-water solvent system and in aqueous caffeine solutions. In both media, the kinetics could be described by a single, second-order process: (formula - see text) with k = 1.8 X 10(6) M-1 s-1 in 40% Me2SO-water [pH 7.4, mu = 0.2 M (NaCl)] and k = 3.9 X 10(7) M-1 s-1 in water [pH 7.4 mu = 0.2 M (NaCl), [caffeine] = 0.025 M]. The reaction shows an ionic strength dependence consistent with a residual 1+ to 2+ charge in the vicinity of the binding region of the protein. The kinetics of the transfer of hemin from albumin to hemopexin (formula - see text) were studied as a function of concentration, ionic strength, pH, and temperature. In experiments conducted at 3 less than or equal to [Alb]0/[Hx]0 less than or equal to 20 where the transfer kinetics are first order, k' = 5 X 10(-3) S-1 at mu = 0.3 M (NaCl), pH 7.1; the reaction is strongly dependent on ionic strength and choice of electrolyte. The addition of imidazole catalyzes this transfer process via a ligand-mediated pathway with k' = 5 X 10(-3) + 21[Im]T2. At [Alb]0/[Hx]0 = 92, the noncatalyzed transfer reaction is second order. From the kinetic analysis of the reaction under these conditions, an estimate is made of the distribution of hemin between the two proteins at concentration levels which are characteristic of serum. The association  of hemin and hemopexin is approximately 30 times faster than that of hemin and albumin, a finding consistent with the recycling function of hemopexin during heme transport to the liver parenchymal cells.
D001798:Blood Proteins / Q000378:metabolism*; D002110:Caffeine; D006418:Heme / Q000031:analogs & derivatives*; D006427:Hemin / Q000378:metabolism*; D006466:Hemopexin / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008433:Mathematics; D011485:Protein Binding; D012709:Serum Albumin / Q000378:metabolism
D001798:Blood Proteins; D012709:Serum Albumin; D002110:Caffeine; D006418:Heme; D006427:Hemin; D006466:Hemopexin