One-step preparation of S-nitrosated human serum albumin with high biological activities.
S-Nitrosated human serum albumin (SNO-HSA) is a large molecular weight nitric oxide carrier in human plasma, and because of its many beneficial effects in different tests, it is currently under investigation as a cytoprotective agent. However, making SNO-HSA preparations is a complicated and time-consuming process. We found that binding of caprylic acid (CA) and N-acetyl-l-tryptophan (N-AcTrp) to defatted mercaptalbumin increased S-nitrosation by S-nitrosoglutathione (GS-NO) by making Cys-34 of HSA more accessible and by protecting it against oxidation, respectively. Fortunately, HSA solutions for clinical use contain high concentrations of CA and N-AcTrp as stabilizers. By making use of that fact it was possible to work-out a fast and simple procedure for producing SNO-HSA: incubation of a commercial HSA formulation with GS-NO for only 1 min results in S-nitrosation of HSA. The biological usefulness of such a preparation was tested in a rat ischemia-reperfusion liver injury model. Although our procedure for making SNO-HSA is fast and straightforward, the cytoprotective effect of the preparation was similar to, or better than, that of a preparation made in a more traditional way. The clinical development of SNO-HSA as a strong cytoprotective agent is under way using this method in collaboration with clinicians and industrial developers.
Ishima
Yu
Y
Department of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmachi, Kumamoto 862-0973, Japan.
Hiroyama
Shuichi
S
Kragh-Hansen
Ulrich
U
Maruyama
Toru
T
Sawa
Tomohiro
T
Akaike
Takaaki
T
Kai
Toshiya
T
Otagiri
Masaki
M
D000818:Animals; D015153:Blotting, Western; D002210:Caprylates / Q000378:metabolism; D002942:Circular Dichroism; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D019610:Cytoprotection / Q000187:drug effects*; D006801:Humans; D008099:Liver / Q000098:blood supply / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism*; D015538:Nitrosation; D009603:Nitroso Compounds / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology* / Q000627:therapeutic use; D010084:Oxidation-Reduction; D051381:Rats; D015427:Reperfusion Injury / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D026422:S-Nitrosoglutathione / Q000737:chemistry; D012709:Serum Albumin / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology* / Q000627:therapeutic use; D000075462:Serum Albumin, Human; D013439:Sulfhydryl Reagents / Q000737:chemistry; D013997:Time Factors; D014364:Tryptophan / Q000031:analogs & derivatives / Q000378:metabolism
D002210:Caprylates; D009603:Nitroso Compounds; C511514:S-nitrosoalbumin, human; D012709:Serum Albumin; D013439:Sulfhydryl Reagents; D009569:Nitric Oxide; C006392:N-acetyltryptophan; D026422:S-Nitrosoglutathione; D014364:Tryptophan; D003545:Cysteine; C031492:octanoic acid; D000075462:Serum Albumin, Human