Human estrogen sulfotransferase (SULT1E1) pharmacogenomics: gene resequencing and functional genomics
Human estrogen sulfotransferase (SULT1E1) pharmacogenomics: gene resequencing and functional genomics
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DOI：
10.1038/sj.bjp.0705369
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发表时间：
2003-08-01
影响因子：
7.3
通讯作者：
Weinshilboum, RM
中科院分区：
文献类型：
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作者：
Adjei, AA;Thomae, BA;Weinshilboum, RM
1雌激素被用作药物，雌激素暴露是乳腺癌等激素依赖性疾病的危险因素。硫酸结合是雌激素代谢的重要途径。磺基转移酶（SULT）SULT 1 E1对10种已知人类SULT亚型中的雌激素和儿茶酚雌激素具有最低的Km值。2我们先前克隆并表征了人类SULT 1 E1 cDNA和基因，作为药物遗传学研究的步骤。在本实验中，我们着手确定SULT 1 E1是否存在常见的、功能显著的遗传多态性。作为第一步，我们使用来自60名非洲裔美国人和60名高加索裔美国人受试者的DNA对8个SULT 1 E1外显子和外显子-内含子剪接点以及5 '侧翼区的部分进行了“重新测序”。3总共观察到23种多态性，22种单核苷酸多态性（SNP）和1种插入缺失。有三个非同义编码SNPs（cSNPs）改变了以下编码的氨基酸：Asp 22 Tyr、Ala 32 Val和Pro253 His。其中，12对SNPs紧密连锁。此外，还鉴定了12种明确的SULT 1 E1单倍型，其中6种是两个研究人群共有的。4在COS-1细胞中瞬时表达含有三种非同义cSNP的构建体，显示SULT 1 E1对Tyr 22和Val 32等位酶的活性显著降低，免疫反应性蛋白水平相应降低。His 253等位酶的活性或免疫反应性蛋白水平没有变化。Val 32等位酶对反应的两种共底物17 β-雌二醇和3 '-磷酸腺苷5'-磷酸硫酸盐的表观Km值与野生型酶的表观Km值没有显著差异，但His 253等位酶的Km值增加了2 - 3倍，Tyr 22等位酶增加了100倍。5这些观察结果提高了SULT 1 E1催化的雌激素硫酸化中遗传决定的变异可能有助于雌激素-β 2受体拮抗剂的病理生理学的可能性。依赖性疾病以及外源性雌激素生物转化的变化。
1 Estrogens are used as drugs and estrogen exposure is a risk factor for hormone-dependent diseases such as breast cancer. Sulfate conjugation is an important pathway for estrogen metabolism. The sulfotransferase (SULT) enzyme SULT1E1 has the lowest K-m values for estrogens and catecholestrogens of the 10 known human SULT isoforms.2 We previously cloned and characterized the human SULT1E1 cDNA and gene as steps toward pharmacogenetic studies. In the present experiments, we set out to determine whether common, functionally significant genetic polymorphisms might exist for SULT1E1. As a first step, we 'resequenced' the eight SULT1E1 exons and exon-intron splice junctions as well as portions of the 5'-flanking region using DNA from 60 African-American and 60 Caucasian-American subjects.3 In all, 23 polymorphisms, 22 single nucleotide polymorphisms (SNPs) and one insertion deletion were observed. There were three nonsynonymous coding SNPs (cSNPs) that altered the following encoded amino acids: Asp22Tyr, Ala32Val and Pro253His. Among these, 12 pairs of SNPs were tightly linked. In addition, 12 unambiguous SULT1E1 haplotypes were identified, including six that were common to both populations studied.4 Transient expression in COS-1 cells of constructs containing the three nonsynonymous cSNPs showed significant decreases in SULT1E1 activity for the Tyr22 and Val32 allozymes, with corresponding decreases in levels of immunoreactive protein. There were no changes in levels of either activity or immunoreactive protein for the His253 allozyme. Apparent K-m values of the Val32 allozyme for the two cosubstrates for the reaction, 17beta-estradiol and 3'-phosphoadenosine 5'-phosphosulfate, were not significantly different from those of the wild-type enzyme, but there was a two- to three-fold increase in K-m values for the His253 allozyme and a greater than five-fold increase for the Tyr22 allozyme.5 These observations raise the possibility that genetically determined variation in SULT1E1-catalyzed estrogen sulfation might contribute to the pathophysiology of estrogen-dependent diseases as well as variation in the biotransformation of exogenously administered estrogens.