HUMAN PLATELET PHENOL SULFOTRANSFERASES

药物药理学的一个主要方面是修饰。 他们的行动和持续时间通过生物转化和 排泄物。硫酸盐化是生物降解的主要途径之一。 药物的生物转化和排泄以及内源性 类固醇和儿茶酚胺等化合物。长期的 本研究的目的是了解硫酸盐化在植物生长中的作用。 人体内药物和内源性底物的新陈代谢。AS 观察其他药物代谢酶，如 谷胱甘肽S转移酶和葡糖醛酸基转移酶， 磺基转移酶，这种酶催化转移 硫酸盐部分为受体化合物，代表一族 不同的同工酶。然而，在人类中， 这些酶的特性和相互关系不是很好 为人所知。 本申请中提出的研究针对的是 蛋白质化学和分子生物学的研究 人酚磺基转移酶。人体血液中的血小板，提供 两种形式的苯酚转移酶(PST)的可及来源 活性、苯酚(P)和儿茶酚胺(M)PST活性。 对血小板PSTs的研究可能提供有价值的见解 硫酸盐在较难接触到的人体组织中的作用。 血小板PST活性也可作为整体的指标。 一种药物通过硫酸盐化作用在体内的代谢。世界银行的目标是 拟议中的研究旨在调查 PSTs在其酶活性、蛋白质水平上 化学和分子生物学。首先，P-和M-PST 我们将对血小板中的活性进行提纯和鉴定。 纯化技术将包括ChromatofocusingR，亲和 层析和特异性抗体识别。P和M 血小板中的PST活性被认为是由 不同的酶，在动力学上也与PST非常相似 在其他组织和器官中的活动表明这两个 酶广泛分布于人体内。第二，具体 将在兔体内产生针对纯化的PSTs的抗体。 这些抗体将被用来鉴定和免疫定量 PST蛋白的存在。第三，编码这两种基因的DNA 酶将被克隆。针对这两种病毒产生的抗体 纯化的PSTs将用于从 Lambda gtll基因文库。因为血小板是去核的， 从人脑和肝脏产生的Lambda gtll文库将被 使用。从这些文库中获得的cDNA将被表征 以便它们可以作为研究该规定的探针， 人PSTs的异质性和结构。

A major aspect of the pharmacology of drugs is the modification of their action and duration of action via biotransformation and excretion. Sulfation is one of the major pathways for the biotransformation and excretion of drugs as well as endogenous compounds such as steroids and catecholamines. The long-term objective of this research is to understand the role of sulfation in the metabolism of drugs and endogenous substrates in humans. As observed for other drug metabolizing enzymes such as the glutathione S-transferases and UDP-glucuronyltransferases, sulfotransferases, the enzymes catalyzing the transfer of the sulfate moiety to an acceptor compound, represent a family of distinct isoenzymes. However, in humans the number, characteristics and relationships of these enzymes are not well known. The research proposed in this application is directed toward investigating the protein chemistry and molecular biology of human phenol sulfotransferases. Human blood platelets, provide an accessible source of two forms of phenol sulfotransferase (PST) activity, the phenol (P) and catecholamine (M) PST activities. Investigation of the platelet PSTs may provide valuable insights into the role of sulfation in less accessible human tissues. Platelet PST activity may also serve as an indicator for the whole body metabolism of a given drug by sulfation. The goals of the proposed research are designed to investigate the biochemistry of the PSTs at the levels of their enzymic activity, protein chemistry, and molecular biology. First, the P- and M-PST activities in platelets will be purified and characterized. Purification techniques will include ChromatofocusingR, affinity chromatography, and specific antibody recognition. The P and M PST activities in platelets are believed to be catalyzed by different enzymes and are also very similar kinetically to PST activities in other tissues and organs indicating that these two enzymes are widely distributed in the body. Second, specific antibodies will be raised in rabbits against the purified PSTs. These antibodies will be used to identify and immunoquantitate the presence of PST proteins. Third, cDNAs encoding the two enzymes will be cloned. Antibodies raised against the two purified PSTs will be used to isolate the respective cDNAs from lambda gtll cDNA libraries. Since platelets are enucleated, lambda gtll libraries generated from human brain and liver will be used. cDNAs obtained from these libraries will be characterized so that they may serve as probes to study the regulation, heterogeneity, and structure of human PSTs.