Oettl Karl, Marsche Gunther
Institute of Physiological Chemistry, Medical University of Graz, Harrachgasse, Graz, Austria.
Methods Enzymol. 2010;474:181-95. doi: 10.1016/S0076-6879(10)74011-8. Epub 2010 Jun 20.
Albumin, the main protein in plasma, is prone to different mechanisms of oxidative modification since extracellular fluids contain only small amounts of antioxidant enzymes. The redox state of cysteine-34 of human albumin defines three fractions which allow to monitor albumin oxidation: mercaptalbumin with a free thiol group, nonmercaptalbumin1 containing a disulfide, and nonmercaptalbumin2 with a sulfinic or sulfonic acid group. These fractions can be separated by HPLC and detected with UV or fluorescence detection. The method is very rugged and only simple sample preparation is needed. It has been used to demonstrate albumin oxidation during exercise, aging, and pathologies like diabetes, liver disease, or renal disease. Problems may arise when high endogenous concentrations of bilirubin or certain drugs are present. The redox state of albumin shows high variability and is hence a valuable tool for the investigation of reversible and irreversible modification of plasma protein.
白蛋白是血浆中的主要蛋白质,由于细胞外液中仅含有少量抗氧化酶,因此易于发生不同的氧化修饰机制。人白蛋白半胱氨酸-34的氧化还原状态定义了三个组分,可用于监测白蛋白氧化:具有游离巯基的巯基白蛋白、含有二硫键的非巯基白蛋白1和具有亚磺酸或磺酸基团的非巯基白蛋白2。这些组分可通过高效液相色谱法分离,并用紫外或荧光检测法进行检测。该方法非常耐用,只需要简单的样品制备。它已被用于证明运动、衰老以及糖尿病、肝病或肾病等病理状态下的白蛋白氧化。当存在高内源性浓度的胆红素或某些药物时,可能会出现问题。白蛋白的氧化还原状态具有高度变异性,因此是研究血浆蛋白可逆和不可逆修饰的有价值工具。