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首页> 美国卫生研究院文献>Scientific Reports >LC-MS/MS quantification of sulfotransferases is better than conventional immunogenic methods in determining human liver SULT activities: implication in precision medicine
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LC-MS/MS quantification of sulfotransferases is better than conventional immunogenic methods in determining human liver SULT activities: implication in precision medicine

机译：LC-MS / MS在确定人肝SULT活性方面优于传统的免疫原性方法对磺基转移酶定量：对精密医学的意义

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This study aims to determine whether enzyme activities are correlated with protein amounts and mRNA expression levels of five major human sulfotransferase (SULT) enzymes in 10 matched pericarcinomatous and hepatocellular carcinoma liver samples. The MRM UHPLC-MS/MS method, Western blot and RT-PCR were used along with SULT activity measurement using probe substrates. The LC-MS/MS method was specific for all five tested SULTs, whereas Western blot was specific for only two isoforms. The activities of SULT1A1, SULT1B1, SULT1E1 and SULT2A1 in 9 of 10 samples showed a significant decrease in tumor tissues relative to matched pericarcinomatous tissues, whereas the activities of SULT1A3 in 7 of 10 samples increased. The turnover numbers of SULTs did not change, except for SULT1A1. A generally high degree of correlations was observed between SULT activities and protein amounts (r2 ≥ 0.59 except one), whereas a low degree of correlations was observed between SULT activities and mRNA expression levels (r2 ≤ 0.48 except one). HCC reduced the SULT activities via impaired protein amounts. LC-MS/MS quantification of SULTs is highly reliable measurement of SULT activities, and may be adopted for implementing precision medicine with respect to drugs mainly metabolized by SULTs in healthy and HCC patients.
机译：这项研究旨在确定在10个匹配的癌旁和肝细胞癌肝样本中，酶活性是否与5种主要人类磺基转移酶（SULT）酶的蛋白质含量和mRNA表达水平相关。使用MRM UHPLC-MS / MS方法，蛋白质印迹和RT-PCR以及使用探针底物的SULT活性测量。 LC-MS / MS方法对所有五个测试的SULT具有特异性，而Western印迹仅对两种同工型具有特异性。 10个样本中有9个样本的SULT1A1，SULT1B1，SULT1E1和SULT2A1活性相对于匹配的癌旁瘤组织显着降低了肿瘤组织，而10个样本中有7个样本的SULT1A3活性却增加了。除了SULT1A1，SULT的周转次数没有变化。在SULT活性和蛋白质含量之间通常观察到高度相关性（r 2 ≥0.59，除了一个），而在SULT活性和mRNA表达水平之间观察到低相关性（r 2 ≤0.48，除了一个）。 HCC通过受损的蛋白质量降低了SULT活性。 LC-MS / MS对SULT的定量是对SULT活性的高度可靠的度量，对于健康和HCC患者中主要由SULT代谢的药物，可用于实施精密医学。

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