Hooper D, Kawamura M, Hoffman B, Kopin I J, Hunyady B, Mezey E, Eisenhofer G
Clinical Neuroscience Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892, USA.
J Chromatogr B Biomed Sci Appl. 1997 Jul 4;694(2):317-24. doi: 10.1016/s0378-4347(97)00166-7.
A nonisotopic assay for tyrosine hydroxylase, with optimized signal-to-noise ratios, enables determination of low levels of enzyme activity in peripheral tissues. DOPA produced by the enzyme is measured using HPLC with electrochemical detection. Increased signal-to-noise ratios are obtained by including in the reaction mixture glycerol for reduction of blank values and dihydropteridine reductase and NADPH for regeneration of the tetrahydropteridine cofactor. With this method, tyrosine hydroxylase activity can be detected in as few as 200 PC12 cells and in peripheral tissues at levels as low as 4.5 fmol/min/mg wet weight. The assay permits activity to be assessed in a variety of peripheral tissues.
一种具有优化信噪比的酪氨酸羟化酶非同位素检测方法，能够测定外周组织中低水平的酶活性。通过高效液相色谱-电化学检测法测定该酶产生的多巴。在反应混合物中加入甘油以降低空白值，加入二氢蝶啶还原酶和烟酰胺腺嘌呤二核苷酸磷酸以再生四氢蝶啶辅因子，从而提高信噪比。采用这种方法，在低至200个PC12细胞以及外周组织中，以低至4.5飞摩尔/分钟/毫克湿重的水平都能检测到酪氨酸羟化酶活性。该检测方法可用于评估多种外周组织中的活性。
