S-亚硝基硫醇的还原测定法：对生物系统中测量的意义。
Reductive assays for S-nitrosothiols: implications for measurements in biological systems.
作者信息
Fang K, Ragsdale N V, Carey R M, MacDonald T, Gaston B
机构信息
Department of Pediatrics, University of Virginia Health Sciences Center, Charlottesville, Virginia, 22908, USA.
出版信息
Biochem Biophys Res Commun. 1998 Nov 27;252(3):535-40. doi: 10.1006/bbrc.1998.9688.
Bioactive SNOs are found in many tissues. We speculated SNOs might be misidentified in conventional assays which reduce NO-3 to NO. S-Nitrosothiols were exposed to saturated VCl3 in HCl, 1% KI in acetic acid, photolysis, or CuCl and CSH in He; NO was measured by chemiluminescence. S-Nitrosothiols were readily detected in VCl3 but not in KI. Reduction in CuCl/cysteine was linear (r2 = 1.0, n = 6), sensitive to 10 pmol, and eliminated by HgCl2; it did not detect NO-2, NO-3, or 3-nitrotyrosine. S-Nitrosothiols represented approximately 2.9% of NOx assayed by VCl3 in human serum, of which <5% were low-mass species. In summary, (i) conventional assays may misidentify NO-3, but not NO-2, as SNOs; and (ii) chemiluminescence/reduction systems may be sensitive and specific as SNO assays. We suggest that assay of the SNO fraction in biological NOx may be more relevant and feasible than is now appreciated.
生物活性硫亚硝基化合物存在于许多组织中。我们推测在将NO₃还原为NO的传统检测方法中，硫亚硝基化合物可能被误识别。硫代亚硝基硫醇分别暴露于HCl中的饱和VCl₃、乙酸中的1% KI、光解或He中的CuCl和CSH中；通过化学发光法测量NO。硫代亚硝基硫醇在VCl₃中很容易被检测到，但在KI中则不然。CuCl/半胱氨酸的还原呈线性（r² = 1.0，n = 6），对10 pmol敏感，并被HgCl₂消除；它不能检测到NO₂、NO₃或3-硝基酪氨酸。硫代亚硝基硫醇约占人血清中用VCl₃检测的NOx的2.9%，其中<5%为低质量物种。总之，（i）传统检测方法可能会将NO₃误识别为硫亚硝基化合物，但不会将NO₂误识别为硫亚硝基化合物；（ii）化学发光/还原系统作为硫亚硝基化合物检测方法可能具有敏感性和特异性。我们认为，检测生物NOx中的硫亚硝基化合物部分可能比目前所认识到的更具相关性和可行性。
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