The determination of homocysteine-thiolactone in biological samples
The determination of homocysteine-thiolactone in biological samples
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DOI：
10.1016/s0003-2697(02)00224-5
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发表时间：
2002-09-01
影响因子：
2.9
通讯作者：
Jakubowski, H
中科院分区：
文献类型：
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作者：
Jakubowski, H
同型半胱氨酸硫内酯是同型半胱氨酸的环状硫酯，由所有细胞类型中的甲硫氨酰-tRNA 合成酶合成。描述了一种测定生物样品中同型半胱氨酸-硫内酯的新方法。该测定涉及通过超滤从大分子中分离同型半胱氨酸-硫内酯。同型半胱氨酸-硫代内酯通过高压液相色谱在反相柱或阳离子交换微孔柱上进一步纯化和定量。通过监测 240 nm 处的吸光度（同型半胱氨酸-硫代内酯的 UV 光谱中的最大值）进行检测和定量。检测灵敏度为5 pmol。该测定已应用于细菌（大肠杆菌和耻垢分枝杆菌）、酿酒酵母、培养的人血管内皮细胞和人血浆。数据支持这样的结论：同型半胱氨酸硫内酯是一种普遍存在的代谢物，其水平与同型半胱氨酸水平直接相关。 (C) 2002 年爱思唯尔科学（美国）。版权所有。
Homocysteine-thiolactone, a cyclic thioester of homocysteine, is synthesized by methionyl-tRNA synthetase in all cell types. A new assay for the determination of homocysteine-thiolactone in biological samples is described. The assay involves separation of homocysteine-thiolactone from macromolecules by ultrafiltration. Homocysteine-thiolactone is further purified and quantified by high-pressure liquid chromatography either on a reverse phase or a cation exchange micro-bore column. The detection and quantitation are obtained by monitoring the absorbance at 240 nm, a maximum in a UV spectrum of homocysteine-thiolactone. The sensitivity of detection is 5 pmol. This assay has been applied to bacteria (Escherichia coli and Mycobacterium smegmatis), the yeast Saccharomyces cerevisiae, cultured human vascular endothelial cells, and human plasma. The data support the conclusion that homocysteine-thiolactone is a ubiquitous metabolite whose levels are directly related to homocysteine levels. (C) 2002 Elsevier Science (USA). All rights reserved.