Carbonic anhydrases are producers of S-nitrosothiols from inorganic nitrite and modulators of soluble guanylyl cyclase in human platelets
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- 作者：Erik Hanff ; Anke Böhmer ; Maximilian Zinke ; Stepan Gambaryan…
- 刊名：Amino Acids
- 出版年：2016
- 出版时间：July 2016
- 年：2016
- 卷：48
- 期：7
- 页码：1695-1706
- 全文大小：1,630 KB
- 刊物类别：Biomedical and Life Sciences
- 刊物主题：Life Sciences
Biochemistry
Analytical Chemistry
Biochemical Engineering
Life Sciences
Proteomics
Neurobiology - 出版者：Springer Wien
- ISSN：1438-2199
- 卷排序：48
文摘
Nitric oxide (NO), S-nitrosoglutathione (GSNO) and S-nitrosocysteine are highly potent signaling molecules, acting both by cGMP-dependent and cGMP-independent mechanisms. The NO metabolite nitrite (NO2−) is a major NO reservoir. Hemoglobin, xanthine oxidoreductase and carbonic anhydrase (CA) have been reported to reduce/convert nitrite to NO. We evaluated the role and the physiological importance of CA for an extra-platelet CA/nitrite/NO/cGMP pathway in human platelets. Authentic NO was analyzed by an NO-sensitive electrode. GSNO and GS15NO were measured by liquid chromatography–tandem mass spectrometry (LC–MS/MS). cGMP was determined by LC–MS/MS or RIA. In reduced glutathione (GSH) containing aqueous buffer (pH 7.4), human and bovine erythrocytic CAII-mediated formation of GSNO from nitrite and GS15NO from 15N-nitrite. In the presence of l-cysteine and GSH, this reaction was accompanied by NO release. Incubation of nitrite with bovine erythrocytic CAII and recombinant soluble guanylyl cyclase resulted in cGMP formation. Upon incubation of nitrite with bovine erythrocytic CAII and washed human platelets, cGMP and P-VASPS239 were formed in the platelets. This study provides the first evidence that extra-platelet nitrite and erythrocytic CAII may modulate platelet function in a cGMP-dependent manner. The new nitrite-dependent CA activity may be a general principle and explain the cardioprotective effects of inorganic nitrite in the vasculature. We propose that nitrous acid (ONOH) is the primary CA-catalyzed reaction product of nitrite.KeywordsCarbonic anhydrasecGMPNitric oxideNitriteS-NitrosothiolsPlatelets