Mechanisms and Functions of Human Sulfotransferases

人类磺基转移酶的机制和功能

• 批准号: 7135375
• 负责人: Guangping Chen
• 金额: $ 25.37万
• 依托单位: OKLAHOMA STATE UNIVERSITY STILLWATER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-22 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7135375
• 关键词: Escherichia coli; X ray crystallography; active sites; cell line; chemical kinetics; computer simulation; drug metabolism; enzyme inhibitors; enzyme mechanism; enzyme substrate complex; human genetic material tag; oxidation reduction reaction; pharmacokinetics; polymerase chain reaction; protein purification; protein sequence; site directed mutagenesis; sulfation; sulfotransferase; thiols; transfection /expression vector; western blottings

DESCRIPTION (provided by applicant): Phase II drug metabolizing enzymes sulfotransferases (SULTs) catalyzed sulfation is important in the regulation of different hormones and the detoxification of drugs and other xenobiotics. Sulfation also leads to bioactivation of procarcinogens leading to toxic effect. The long-term goal of this research project is to understand human SULT biological functions and to investigate their relevance to human health under physiological and pathological conditions. Specific aims in this proposal are as follows: 1. To investigate mechanisms of catalysis, substrate inhibition, and product inhibition/activation of human SULTs. The proposed bypass ordered mechanism and related alternative mechanisms will be investigated using kinetic analysis, isotope exchange, sulfated active site amino acid residue identification, and site-directed mutagenesis. The information will have important implications for the prediction of biotransformation pathways. 2. To investigate the effect of sulfated drugs on human SULT catalytic activities. The inhibition and activation effect of clinically important drug sulfates on catalytic activities of human SULTs will be investigated. E. coli expressed and purified human SULTs; human intestinal cytosols; and human Hep G2 and Caco-2 cells will be used for these investigations. The effect of clinical drugs on human SULT activities may interfere SULT normal biological functions in hormone regulation and xenobiotic detoxification. 3. To define oxidative regulation mechanisms of human SULT1E1. Oxidative regulation of human SULT1E1 in Hep G2 and Caco-2 cells and redox thiol regulation mechanisms of purified human SULT1E1 will be investigated using enzyme assay, Western blot, RT-PCR, amino acid modification, site-directed mutagenesis, kinetic analysis, crystal structure analysis, and computer modeling methods. Knowledge on oxidative regulation of certain human SULT is important in understanding the ability of SULT functioning under physiological and pathological conditions. This proposal studies human SULTs. These studies will be significant in understanding SULT biological functions including hormone regulation, drug metabolism, xenobiotic detoxification and procarcinogen bioactivation. The knowledge will be important in understanding drug side effect, drug-drug interaction, drug development, and the potential roles SULTs play in cancer prevention and causation.

描述（由申请人提供）：II期药物代谢酶硫代转移酶（SULTS）催化硫酸化在调节不同激素以及药物和其他异种生物学的排毒中很重要。硫酸化还导致促促促促炎性的生物活化导致毒性作用。该研究项目的长期目标是了解人类的苏尔特生物学功能，并在生理和病理条件下研究其与人类健康的相关性。该提案中的具体目的如下：1。研究人类苏尔特的催化机制，底物抑制和产物抑制/激活。拟议的旁路有序机制和相关的替代机制将使用动力学分析，同位素交换，硫酸化活性位点氨基酸残基鉴定和定向诱变。该信息将对预测生物转化途径具有重要意义。 2。研究硫酸药物对人类苏尔特催化活性的影响。将研究临床上重要的药物硫酸盐对人类苏尔特催化活性的抑制和激活作用。大肠杆菌表达和纯净的人类宗教；人类肠道胞质；这些研究将使用人HEP G2和CACO-2细胞。临床药物对人类苏尔特活性的影响可能会干扰激素调节和异种生物学解毒的正常生物学功能。 3。定义人类Sult1E1的氧化调控机制。将使用酶测定，Western Blot，RT-PCR，氨基酸修饰，位置定向的诱变，动力学分析，晶体结构分析和计算机建模方法来研究HEP G2和CACO-2细胞和CACO-2细胞中人类Sult1E1的氧化调节以及氧化硫代硫醇调节机制。关于某些人类苏尔特的氧化调节知识对于理解生理和病理条件下的苏尔特功能的能力很重要。该建议研究人类的宗教。这些研究将在理解包括激素调节，药物代谢，异种源解毒和procarcinogen生物活化在内的苏尔特生物学功能方面具有重要意义。知识对于理解药物副作用，药物 - 药物相互作用，药物开发以及苏尔特在预防癌症和因果关系中的潜在作用将很重要。