Human serum albumin incorporating tetrakis(o-pivalamido)phenylporphinatoiron(II) derivative as a totally synthetic O-2-carrying hemoprotein

2-[8-{N-(2-Methylimidazolyl)}octanoyloxymethyl]-5,10,15,20-tetrakis(o-pival amido)phenylphorin-atoiron(II)s (FePs) were incorporated into hydrophobic c avities of recombinant human serum albumin (rHSA), providing a totally synt hetic O-2-carrying hemoprotein (rHSA-FeP). An rHSA host absorbs maximally e ight FeP molecules. Solution properties of the obtained albumin hybrid [[rH SA] = 5 wt%; FeP/HSA = 1-8 (mol/mol)] are almost identical to those of the rHSA itself; the specific gravity is 1.013 and the viscosity is 1.1 cP. Cir cular dichroism spectroscopy and isoelectric focusing measurement revealed that the second-order structure and surface charge distribution of rHSA wer e always constant independent of the binding numbers of FeP. Hydrophobic in teraction is probably a major molecular force of the incorporation of this synthetic heme. rHSA-FeP can bind and release dioxygen reversibly under phy siological conditions (in aqueous media, pH 7.3, 37 degrees C) like hemoglo bin and myoglobin. Its O-2-coordination structure was evaluated by resonanc e Raman spectroscopy. The O-2 rebinding after the laser flash photolysis sh owed three-phases decay, which were analyzed by triple-exponential kinetics . The O-2-binding affinity and O-2-association and -dissociation rate const ants of rHSA-FeP satisfy the initial clinical requirements for O-2 infusion as a red cell substitute.