ANALYSIS OF BIOLOGICAL THIOLS - DERIVATIZATION WITH MONOBROMOTRIMETHYLAMMONIOBIMANE AND CHARACTERIZATION BY ELECTROPHORESIS AND CHROMATOGRAPHY
ANALYSIS OF BIOLOGICAL THIOLS - DERIVATIZATION WITH MONOBROMOTRIMETHYLAMMONIOBIMANE AND CHARACTERIZATION BY ELECTROPHORESIS AND CHROMATOGRAPHY
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DOI：
10.1016/0003-2697(80)90483-2
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发表时间：
1980-01-01
影响因子：
2.9
通讯作者：
KOSOWER, EM
中科院分区：
文献类型：
--
作者：
FAHEY, RC;NEWTON, GL;KOSOWER, EM
介绍了一种基于生物硫醇与一溴三甲基氨二亚胺（qBBr）反应生成荧光衍生物的新分析方法。衍生物通过色谱法和在纤维素TLC板上电泳分离。二维映射的使用使得有可能区分各种各样的生物硫醇，包括N-乙酰半胱氨酸、CoA、半胱氨酸、半胱氨酰甘氨酸、半胱胺、麦角硫因、谷胱甘肽、γ-谷氨酰胺、γ-谷氨酰胺和γ-谷氨酰胺。谷氨酰半胱氨酸、高半胱氨酸、巯基嘧啶、泛酰巯基乙胺、4“-磷酸泛酰巯基乙胺、硫代硫酸盐和硫尿嘧啶。对于生物样品的应用，通过与汞琼脂糖凝胶结合从粗提取物中分离硫醇。用二硫苏糖醇从凝胶中除去后，用qBBr衍生化硫醇。通过显示谷胱甘肽是人红细胞中的主要硫醇，谷胱甘肽和麦角硫因是粗糙脉孢菌分生孢子中的主要硫醇，以及蜡状芽孢杆菌营养细胞缺乏谷胱甘肽但含有显著量的半胱氨酸、泛酰巯基乙胺和未鉴定的硫醇来测试所述方法。
A new method for analysis of biological thiols based upon their conversion to fluorescent derivatives by reaction with monobromotrimethylammoniobimane (qBBr) is described. The derivatives are separated by chromatography and by electrophoresis on cellulose TLC plates. The use of 2-dimensional mapping makes it possible to differentiate between a wide variety of biological thiols including N-acetylcysteine, CoA, cysteine, cysteinylglycine, cysteamine, ergothioneine, glutathione, .gamma.-glutamylcysteine, homocysteine, mercaptopyrimidine, pantetheine, 4''-phosphopantetheine, thiosulfate and thiouracil. For applications to biological samples, thiols were isolated from crude extracts by binding to a mercuriagarose gel. Following removal from the gel with dithiothreitol, the thiols were derivatized with qBBr. The methods were tested by showing that glutathione is the major thiol in human red blood cells, that glutathione and ergothioneine are the major thiols in Neurospora crassa conidia and that Bacillus cereus vegetative cells lack glutathione but contain cysteine, pantetheine, and an unidentified thiol in significant amounts.