Roer R D, Halbrook K E, Shafer T H
Department of Biological Sciences, University of North Carolina at Wilmington, 601 South College Road, Wilmington, NC 28403-3297, USA.
Comp Biochem Physiol B Biochem Mol Biol. 2001 Apr;128(4):683-90. doi: 10.1016/s1096-4959(00)00363-8.
We have previously demonstrated a marked change in sugar moieties of glycoproteins of the cuticle of the blue crab, Callinectes sapidus, between 0.5 and 3 h post-ecdysis. The present study has identified a glycosidase that appears in the cuticle during the early post-ecdysial hours. The enzyme has affinities for p-nitrophenyl derivatives of both N-acetylglucosamine and N-acetylgalactosamine. Both activities are competitively inhibited by chitobiose, suggesting that the enzyme could be a N-acetylhexosaminidase (EC 3.2.1.52). Atypical of N-acetylhexosaminidases described to date, this enzyme has a pH optimum of 7.0. The enzyme activity is high during the post-ecdysial period coincident with the changes in glycoprotein profiles observed in vivo. Partial purification of the enzyme has been accomplished by Sephacryl size-exclusion chromatography followed by concanavalin A affinity chromatography.
我们之前已经证明,在蓝蟹(Callinectes sapidus)蜕皮后0.5至3小时之间,其表皮糖蛋白的糖基部分发生了显著变化。本研究鉴定出一种在蜕皮后早期出现在表皮中的糖苷酶。该酶对N-乙酰葡糖胺和N-乙酰半乳糖胺的对硝基苯基衍生物均有亲和力。两种活性均受到壳二糖的竞争性抑制,这表明该酶可能是一种N-乙酰己糖胺酶(EC 3.2.1.52)。与迄今描述的N-乙酰己糖胺酶不同,该酶的最适pH为7.0。在蜕皮后期间,该酶活性很高,这与体内观察到的糖蛋白谱变化一致。通过Sephacryl尺寸排阻色谱,然后是伴刀豆球蛋白A亲和色谱,已完成该酶的部分纯化。