Oxidation of Arg-410 promotes the elimination of human serum albumin.
The effect of the oxidation of amino acid residues on albumin on its in vivo elimination was investigated using mutants and oxidized HSAs. The single-residue mutants (H146A, K199A, W214A, R218H, R410A, Y411A) and oxidized HSAs were produced by the recombinant DNA techniques and incubation with a metal ion-catalyzed oxidation (MCO) system for 12, 24, 48 or 72 h. Pharmacokinetics were evaluated in mice after labeling with 111In. Structural and functional properties were examined by several spectroscopic techniques. Time-dependent increase in carbonyl group content resulted in increase in the liver clearance of oxidized HSAs. Slight decreases in alpha-helical content as the result of oxidation was induced by the increases in accessible hydrophobic areas and the net negative charge on the HSA molecule. No significant change in the pharmacokinetics and structural properties was observed for the W214A, R218H and Y411A mutants, but the properties for the H146A, K199A and R410A mutants were affected (extent of effect: R410A > K199A > H146A). The liver clearance of these proteins is closely correlated to hydrophobicity (r = 0.929, P < 0.01) and the net charge of the proteins (r=0.930, P < 0.01). The rate of elimination of HSA is closely related to the hydrophobicity and net charge of the molecule. Further, the R410A mutants had a short half-life and structure similar to oxidized HSA after oxidation. Therefore, the modification of Arg-410 via oxidative stress may promote the elimination of HSA.
Department of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto 862-0973, Japan.
Yasunori
Y
Iwao
Makoto
M
Anraku
Keishi
K
Yamasaki
Ulrich
U
Kragh-Hansen
Keiichi
K
Kawai
Toru
T
Maruyama
Masaki
M
Otagiri
D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000737:chemistry*; D006207:Half-Life; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007660:Ketoprofen / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008297:Male; D051379:Mice; D010084:Oxidation-Reduction; D017354:Point Mutation; D011487:Protein Conformation; D012709:Serum Albumin / Q000235:genetics / Q000378:metabolism* / Q000493:pharmacokinetics; D014859:Warfarin / Q000378:metabolism
D012709:Serum Albumin; D014859:Warfarin; D007660:Ketoprofen; D001120:Arginine