Rivoal J., Hanson A. D.
Institut de Recherche en Biologie Vegetale de l'Universite de Montreal, 4101 Rue Sherbrooke Est, Montreal, Quebec, H1X 2B2, Canada.
Plant Physiol. 1994 Nov;106(3):1187-1193. doi: 10.1104/pp.106.3.1187.
Choline-O-sulfate is a compatible osmolyte accumulated under saline conditions by members of the halophytic genus Limonium and other Plumbaginaceae. A choline sulfotransferase (EC 2.8.2.6) responsible for the formation of choline-O-sulfate was characterized in Limonium species. A simple radiometric assay was developed in which [14C]choline was used as substrate, and the h [14C]choline-O-sulfate product was isolated by ion-exchange chromatography. The choline sulfotransferase activity was soluble, required 3[prime]-phosphoadenosine-5[prime]-phosphosulfate as the sulfate donor, and showed a pH optimum at 9.0. Apparent Km values were 25 [mu]M for choline and 5.5 [mu]M for 3[prime]-phosphoadenosine-5[prime]-phosphosulfate. Choline sulfotransferase activity was detected in various Limonium species but was very low or absent from species that do not accumulate choline-O-sulfate. In roots and leaves of Limonium perezii, the activity was increased at least 4-fold by salinization with 40% (v/v) artificial sea water. Choline sulfotransferase activity was also induced in cell cultures of L. perezii following salt shock with 20% (v/v) artificial sea water or osmotic shock with 19% (w/v) polyethylene glycol 6000. Labeling experiments with [14C]choline confirmed that the enzyme induced in cell cultures was active in vivo.
硫酸胆碱-O-酯是一种相容性渗透剂,盐生补血草属植物及其他蓝雪科植物在盐渍条件下会积累该物质。在补血草属植物中鉴定出了一种负责硫酸胆碱-O-酯形成的胆碱磺基转移酶(EC 2.8.2.6)。开发了一种简单的放射性测定法,其中使用[14C]胆碱作为底物,并通过离子交换色谱法分离出[14C]硫酸胆碱-O-酯产物。胆碱磺基转移酶活性是可溶的,需要3'-磷酸腺苷-5'-磷酸硫酸作为硫酸盐供体,并且在pH 9.0时表现出最佳活性。胆碱的表观Km值为25μM,3'-磷酸腺苷-5'-磷酸硫酸的表观Km值为5.5μM。在各种补血草属植物中均检测到胆碱磺基转移酶活性,但在不积累硫酸胆碱-O-酯的物种中该活性非常低或不存在。在佩氏补血草的根和叶中,用40%(v/v)人工海水盐渍可使活性至少提高4倍。在用20%(v/v)人工海水进行盐胁迫或用19%(w/v)聚乙二醇6000进行渗透胁迫后,佩氏补血草的细胞培养物中也诱导出了胆碱磺基转移酶活性。用[14C]胆碱进行的标记实验证实,细胞培养物中诱导产生的酶在体内具有活性。