NITROTYROSINE AS A NEW MARKER FOR ENDOGENOUS NITROSATION AND NITRATION OF PROTEINS
NITROTYROSINE AS A NEW MARKER FOR ENDOGENOUS NITROSATION AND NITRATION OF PROTEINS
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DOI：
10.1016/0278-6915(90)90173-k
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发表时间：
1990-09-01
影响因子：
4.3
通讯作者：
BARTSCH, H
中科院分区：
文献类型：
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作者：
OHSHIMA, H;FRIESEN, M;BARTSCH, H
组织或血液蛋白中的3-硝基酪氨酸（NTYR）被评估为外源性和内源性亚硝化或硝化剂的可能暴露标记。建立了气相色谱-热能分析仪（GC-TEA）分析NTYR的灵敏选择性方法。利用这种方法，进行了一系列动力学研究。发现游离和蛋白结合的酪氨酸残基很容易与硝化/亚硝化剂反应生成NTYR。注射不同剂量（0.5-2.5 μ mol/鼠）的四硝基甲烷24小时后，大鼠血浆蛋白和血红蛋白中NTYR的形成呈剂量依赖性增加。大鼠口服NTYR的主要尿代谢产物为3-硝基-4-羟基苯基乙酸（NHPA）和3-硝基-4-羟基苯基乳酸（NHPL）。口服NTYR （100 μ g/大鼠）的44%和5%分别以NHPA和NHPL的形式排出体外。11份24小时人尿经乙酸乙酯萃取和高效液相色谱纯化后，采用气相色谱- tea分析NHPA含量：浓度范围为0 ~ 7.9 mu-g/24小时，平均+/- SD为2.8 +/- 2.3 (n = 11)（检出限为0.2 mu-g/升）。尿中蛋白质或其代谢物中的NTYR作为内源性亚硝化和硝化的一种新的/额外的标记物，可以很容易地被GC-TEA分析。
3-Nitrotyrosine (NTYR) in tissue or blood proteins was evaluated as a possible exposure marker for exogenous and endogenous nitrosating or nitrating agents. A sensitive and selective method for analysing NTYR by gas chromatography with a thermal energy analyser (GC-TEA) was developed. Using this method, a number of kinetic studies were carried out. It was found that free and protein-bound tyrosine residues easily react with nitrating/nitrosating agents to yield NTYR. NTYR formation in vivo showed a dose-dependent increase in NTYR in both plasma proteins and haemoglobin obtained from rats 24 hr after ip injection of various doses (0.5-2.5 mu-mol/rat) of tetranitromethane. Major urinary metabolites of NTYR, given orally to rats, were isolated and identified as 3-nitro-4-hydroxyphenylacetic acid (NHPA) and 3-nitro-4-hydroxyphenyllactic acid (NHPL). About 44% and 5% of the oral dose of NTYR (100 mu-g/rat) was excreted as NHPA and NHPL, respectively. Eleven 24-hr human urine samples were analysed for NHPA by GC-TEA after ethyl acetate extraction and HPLC purification: quantities ranging from 0 to 7.9 mu-g/24 hr, mean +/- SD 2.8 +/- 2.3 (n = 11) were detected (detection limit 0.2 mu-g/litre). NTYR in proteins or its metabolites in urine can be readily analyzed by GC-TEA as a new/additional marker for endogenous nitrosation and nitration.