S-nitrosylation of human variant albumin Liprizzi (R410C) confers potent antibacterial and cytoprotective properties

Yu Ishima, Tomohiro Sawa, Ulrich Kragh-Hansen, Yoichi Miyamoto, Sadaharu Matsushita, Takaaki Akaike, Masaki Otagiri

Research output: Contribution to journalArticlepeer-review

49 Citations (Scopus)

Abstract

The S-nitrosylated forms of certain proteins such as albumin have been thought to be circulating endogenous reservoirs of nitric oxide (NO) and may have potential as NO donors in therapeutic applications. In this study, we investigated the characteristics of R410C, a genetic variant of human serum albumin with two free thiols at positions 34 (Cys-34) and 410 (Cys-410), as a NO carrier via S-nitroso formation. A biotin switch assay revealed that Cys-410 was more rapidly and efficiently nitrosylated than was Cys-34. Nitrosylation of Cys-410 introduced only small conformational changes in the protein, which were detected by far-UV circular dichroism but not by near-UV circular dichroism. In addition, both native R410C and S-nitrosylated R410C did not induce molecular heterogeneity through oligomerization. S-Nitrosylated R410C exhibited strong antibacterial activity against Salmonella typhimurium in vitro and suppressed apoptosis of U937 human promonocytic cells induced by Fas ligand. In a rat ischemia-reperfusion liver injury model, S-nitrosylated R410C treatment significantly reduced liver damage, as indicated by markedly decreased release of liver enzymes (aspartate aminotransferase and alanine aminotransferase). Pharmacokinetic analyses indicated retention of the S-nitroso moiety of S-nitrosylated R410C in circulation after i.v. injection, with an approximate half-life of 20.4 min in the mouse. These data suggest that R410C can be a useful NO carrier and can be regarded as a new class of S-nitrosylated proteins possessing antibacterial and cytoprotective properties with a circulation time sufficient for in vivo biological activity.

Original languageEnglish
Pages (from-to)969-977
Number of pages9
JournalJournal of Pharmacology and Experimental Therapeutics
Volume320
Issue number3
DOIs
Publication statusPublished - 2007 Mar

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology

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