Whittemore R M, Pearce L B, Roth J A
Arch Biochem Biophys. 1986 Sep;249(2):464-71. doi: 10.1016/0003-9861(86)90023-8.
The identification of three forms of phenol sulfotransferase (PST) in human brain and the subsequent purification and kinetic characterization of a phenol-sulfating form of the enzyme are described. Two forms of PST which were capable of conjugating phenol and a third form which sulfated dopamine were resolved from one another using DEAE-cellulose chromatography. One of the phenol-sulfating forms (P1-PST) was subsequently purified on Affi-Gel blue and Sephacryl S-200, giving a final purification of almost 390-fold, with an overall yield of approximately 5%. The purified enzyme was sensitive to NaCl and showed an optimum for phenol conjugation at pH 8.5. Kinetic analysis demonstrated that sulfation by P1-PST proceeds via a sequential ordered, bi-substrate reaction mechanism, where 3'-phosphoadenosine-5'-phosphosulfate (PAPS) is the leading substrate. The true Km and Kia values for PAPS were both 0.35 microM, while the true Km value for phenol was 2.8 microM.
本文描述了在人脑中鉴定出三种形式的酚磺基转移酶（PST），以及随后对该酶的一种酚硫酸化形式进行纯化和动力学表征的过程。使用DEAE - 纤维素色谱法将两种能够结合酚的PST形式与第三种硫酸化多巴胺的形式彼此分离。随后，其中一种酚硫酸化形式（P1 - PST）在Affi - Gel蓝和Sephacryl S - 200上进行纯化，最终纯化倍数接近390倍，总产率约为5%。纯化后的酶对NaCl敏感，在pH 8.5时对酚结合表现出最佳活性。动力学分析表明，P1 - PST的硫酸化反应通过顺序有序的双底物反应机制进行，其中3'-磷酸腺苷-5'-磷酸硫酸酯（PAPS）是主要底物。PAPS的真实Km和Kia值均为0.35 microM，而酚的真实Km值为2.8 microM。
