Dityrosine as a product of oxidative stress and fluorescent probe
Dityrosine as a product of oxidative stress and fluorescent probe
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DOI：
10.1007/s00726-003-0014-z
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发表时间：
2003-12-01
期刊：
影响因子：
3.5
通讯作者：
Anderson, SR
中科院分区：
文献类型：
--
作者：
Malencik, DA;Anderson, SR
二核苷酸可以是蛋白质结构的天然成分，也可以是环境胁迫的产物，也可以是体外蛋白质修饰的产物。它既是一种交联剂，也是一种报告交联型蛋白质分子结构和功能信息的荧光探针。不同的反应会产生酪氨酸基，而酪氨酸基又会偶联生成二硫代赖氨酸。蛋白质水解物中二氢酪氨酸的鉴定和定量通常采用反相高压液相色谱(RP-HPLC)或气相色谱。用达比西林衍生的蛋白质水解物的反相高效液相色谱给出了完整的氨基酸分析，其中包括二叔丁氨酸和3-硝基酪氨酸。钙调蛋白含有一对酪氨酸残基，它既经历了光激活，也经历了酶催化的二氨基酪氨酸的形成。偏振测量，利用二硫代赖氨酸的本征荧光，以及催化活性的测定，显示了不同的分子间和分子内交联模式如何影响钙调蛋白与钙离子和酶的相互作用。
Dityrosine can be a natural component of protein structure, a product of environmental stress, or a product of in vitro protein modification. It is both a cross-link and a fluorescent probe that reports structural and functional information on the cross-linked protein molecule. Diverse reactions produce tyrosyl radicals, which in turn may couple to yield dityrosine. Identification and quantitation of dityrosine in protein hydrolysates usually employs reversed phase high pressure liquid chromatography (RP-HPLC) or gas chromatography. RP-HPLC of protein hydrolysates that have been derivatized with dabsyl chloride gives a complete amino acid analysis that includes dityrosine and 3-nitrotyrosine. Calmodulin, which contains a single pair of tyrosyl residues, undergoes both photoactivated and enzyme-catalyzed dityrosine formation. Polarization measurements, employing the intrinsic fluorescence of dityrosine, and catalytic activity determinations show how different patterns of inter- and intramolecular cross-linking affect the interactions of calmodulin with Ca2+ and enzymes.