Purification and characterization of an ethanol-induced UDP-glucuronosyltransferase.
Purification and characterization of an ethanol-induced UDP-glucuronosyltransferase.
复制标题
乙醇诱导的 UDP-葡萄糖醛酸基转移酶的纯化和表征。
DOI：
10.1016/0003-9861(89)90157-4
复制
发表时间：
1989
影响因子：
3.9
通讯作者：
Yost,GS
中科院分区：
文献类型：
--
作者：
Hutabarat,RM;Yost,GS
摘要从乙醇诱导的雄性新西兰白色兔肝蛋白中分离到一种UDP-葡萄糖醛酸转移酶（GT）。动物用含10%乙醇的饮用水预处理2周。通过阴离子交换和亲和层析纯化GT酶，并通过十二烷基硫酸钠-聚丙烯酰胺平板凝胶电泳显示为均相的。乙醇诱导的UDP-葡萄糖醛酸基转移酶的分子量被确定为57，000 Da。通过HPLC分析，乙醇诱导的GT和来自对照兔肝脏的类似纯化的GT的胰蛋白酶解物似乎是不同的，即使酶的分子量是不可区分的。两种蛋白质的氨基酸组成有6种氨基酸不同。乙醇诱导的GT酶以1-萘酚和吗啡为底物的表观Km值分别为43和109 μ m。这些底物的乙醇诱导的GT酶的表观Vmax值分别为83和4.6 nmol/min/mg蛋白质。增加的催化效率，表观V最大Km为1-萘酚和吗啡，乙醇诱导的同工酶相比，控制同工酶活性分别为2.0和2.4倍。在绵羊中提出的兔乙醇诱导GT的多克隆抗体表现出520倍的选择性沉淀的乙醇诱导的蛋白质，而不是控制蛋白质。这些结果表明，产生一种独特的UDP-葡萄糖醛酸基转移酶同工酶，这是产生于兔作为一个结果，长期乙醇暴露。
Abstract A UDP-glucuronosyltransferase (GT) enzyme was isolated from ethanol-induced male New Zealand white rabbit hepatic protein. The animals were pretreated for 2 weeks with 10% ethanol in their drinking water. The GT enzyme was purified by anion-exchange and affinity chromatography and was shown to be homogeneous by sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis. The molecular mass of the ethanol-induced UDP-glucuronosyltransferase was determined to be 57,000 Da. Tryptic digests of the ethanol-induced GT and a similarly purified GT from control rabbit liver appeared to be different by HPLC analysis, even though the molecular masses of the enzymes were indistinguishable. Amino acid compositions of the two proteins were different for six amino acids. The apparent K m values for the ethanol-induced GT enzyme for 1-naphthol and morphine as substrates were 43 and 109 μ m, respectively. The apparent V max values for the ethanol-induced GT enzyme for these substrates were 83 and 4.6 nmol/min/mg protein. The increases in catalytic efficiencies, apparent V max K m for 1-naphthol and for morphine, for the ethanol-induced isozyme compared to the control isozyme activities were 2.0-and 2.4-fold. A polyclonal antibody raised in sheep to the rabbit ethanol-induced GT demonstrated a 520-fold selectivity for precipitation of the ethanol-induced protein rather than the control protein. These results demonstrate the production of an unique isozyme of UDP-glucuronosyltransferase that is produced in rabbits as a result of chronic ethanol exposure.
登录
查看更多内容
编码两种形式 UDP 葡萄糖醛酸基转移酶的小鼠肝脏 mRNA 的无细胞翻译。
影响因子：
3.9
作者：
P. Mackenzie;F. Gonzalez;I. Owens
通讯作者：
I. Owens
葡萄糖醛酸基转移酶：药理学家可以从分子生物学和细胞酶学中期待哪些进展？
影响因子：
5.8
作者：
G. Siest;B. Antoine;S. Fournel;J. Magdalou;J. Thomassin
通讯作者：
J. Thomassin
兔肝雌酮和对硝基苯酚 UDP-葡萄糖醛酸转移酶特性的纯化。
影响因子：
3.9
作者：
Tukey,RH;Tephly,TR
通讯作者：
Tephly,TR
苯巴比妥和 3-甲基胆蒽治疗大鼠的肝微粒体和分离肝细胞中各种药物的葡萄糖苷酸形成。
影响因子：
5.8
作者：
D. Ullrich;K. Bock
通讯作者：
K. Bock
乙醇摄入对小鼠肝毒性剂量的扑热息痛代谢的影响。
DOI：
10.3109/00498258609043556
发表时间：
1986
期刊：
Xenobiotica; the fate of foreign compounds in biological systems
影响因子：
--
作者：
J. Tredger;H. Smith;R. B. Read;R. Williams
通讯作者：
R. Williams