Rodent tissue alpha-L-fucosidases: analysis of brain and spleen isoforms and characterization of the purified hamster liver enzyme.
Rodent tissue alpha-L-fucosidases: analysis of brain and spleen isoforms and characterization of the purified hamster liver enzyme.
复制标题
啮齿动物组织 α-L-岩藻糖苷酶：脑和脾同工型分析以及纯化仓鼠肝酶的表征。
DOI：
10.1016/0305-0491(93)90083-h
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发表时间：
1993
期刊：
影响因子：
--
通讯作者：
Alhadeff,JA
中科院分区：
文献类型：
--
作者：
Shoarinejad,F;Alhadeff,JA
1.四种啮齿动物（大鼠、小鼠、豚鼠、仓鼠）的脑和脾α-L-岩藻糖苷酶的等电聚焦表明，只有小鼠和仓鼠组织含有具有显着活性高于pI 7.0的亚型。 2. 通过琼脂糖-ε-氨基己酰基-岩藻糖胺亲和层析，仓鼠肝脏 α-L-岩藻糖苷酶纯化约 57,000 倍，产率 80%（最终比活性为 24,700 nmol/min/mg 蛋白质）。 3. 仓鼠肝脏α-L-岩藻糖苷酶的SDS-PAGE分析表明存在一到两个间隔紧密的56和60 kDa亚基。蛋白质印迹分析表明，仓鼠酶可被多克隆抗体识别，但不能被单克隆抗体识别（两种抗体均针对人肝脏 α-L-岩藻糖苷酶制备）。 4. 仓鼠肝脏α-L-岩藻糖苷酶的pH-活性曲线较宽，最佳值集中在pH 6.8附近，在pH 5.5至7.5之间具有高活性。
1. Isoelectric focusing of brain and spleen alpha-L-fucosidases from four rodents (rat, mouse, guinea-pig, hamster) indicated that only mouse and hamster tissues contained isoforms with significant amounts of activity above pI 7.0. 2. Hamster liver alpha-L-fucosidase was purified approximately 57,000-fold in 80% yield (to a final specific activity of 24,700 nmol/min/mg protein) by affinity chromatography on agarose-epsilon-aminocaproyl-fucosamine. 3. SDS-PAGE analysis of hamster liver alpha-L-fucosidase indicated the presence of one to two closely-spaced subunits at 56 and 60 kDa. Western blotting analysis indicated the hamster enzyme was recognized by polyclonal antibodies but not by a monoclonal antibody (both antibodies prepared against human liver alpha-L-fucosidase). 4. The pH-activity curve of hamster liver alpha-L-fucosidase is broad with an optimum centered around pH 6.8 and with high activity between pH 5.5 and 7.5.