Analysis of the subunits, isoforms and substrate specificity of mouse liver alpha-L-fucosidase.
Analysis of the subunits, isoforms and substrate specificity of mouse liver alpha-L-fucosidase.
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小鼠肝脏 α-L-岩藻糖苷酶的亚基、亚型和底物特异性分析。
DOI：
10.1016/0305-0491(93)90179-9
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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：
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作者：
Shoarinejad,F;Johnson,SW;Alhadeff,JA
1. SDS-PAGE表明存在小鼠岩藻糖苷酶的两个主要蛋白条带（57和62 kDa），Western印迹表明这两个条带与针对人肝岩藻糖苷酶产生的多克隆抗体（PAb）和/或单克隆抗体（MAb）具有免疫反应性。凝集素SNA和GNA识别两种小鼠蛋白条带，表明两种亚基都是糖基化的并且含有唾液酸残基。2.聚丙烯酰胺凝胶等电聚焦（PAG-IEF）表明，小鼠肝岩藻糖苷酶含有至少7种异构体，其中3种异构体的pI 6.0以上，这在人肝岩藻糖苷酶中未检测到。印迹表明PAb识别7种小鼠岩藻糖苷酶同种型（pIs 3.6-6.8），而4种MAb似乎不识别任何小鼠同种型。3.通过SDS-PAGE研究小鼠α-L-岩藻糖苷酶的分离的同种型的亚基组成。在每种亚型中发现1 - 2条间隔紧密的蛋白质条带，相对于更中性的亚型，酸性更强的亚型中高M（r）条带的相对量有增加的趋势。4.人类和小鼠肝脏α-L-岩藻糖苷酶以相当的速率从寡糖和糖脂中水解L-Fuc，除了神经节苷脂Fuc-GMI被人类α-L-岩藻糖苷酶水解而不是被小鼠α-L-岩藻糖苷酶水解。
1. SDS-PAGE indicates the presence of two major protein bands (57 and 62 kDa) for mouse fucosidase and Western blotting indicates that both bands are immunoreactive with polyclonal antibodies (PAbs) and/or monoclonal antibodies (MAbs) raised against human liver fucosidase. The lectins SNA and GNA recognized both mouse protein bands, indicating that both subunits are glycosylated and contain sialic acid residues. 2. Polyacrylamide gel-isoelectric focusing (PAG-IEF) indicated that mouse liver fucosidase contains at least seven isoforms, with three isoforms above pI 6.0, which were not detected in human liver fucosidase. Blotting indicates that the PAbs recognized seven mouse fucosidase isoforms (pIs 3.6-6.8) whereas the four MAbs did not appear to recognize any of the mouse isoforms. 3. The subunit composition of the separated isoforms of mouse alpha-L-fucosidase was investigated by SDS-PAGE. One-to-two closely-spaced protein bands are found in each isoform with a trend of increasing relative amounts of the high-M (r) band in the more acidic isoforms relative to the more neutral isoforms. 4. Human and mouse liver alpha-L-fucosidases hydrolyze L-Fuc from oligosaccharides and glycolipids at comparable rates, with the exception of ganglioside Fuc-GMI which was hydrolyzed by human, but not by mouse, alpha-L-fucosidase.