The macromolecular composition of Xenopus laevis egg jelly coat.
The macromolecular composition of Xenopus laevis egg jelly coat.
复制标题
非洲爪蟾卵果冻外衣的大分子成分。
DOI：
10.1021/bi00685a010
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发表时间：
1975
期刊：
影响因子：
2.9
通讯作者：
Jerry L. Hedrick
中科院分区：
文献类型：
--
作者：
Edward C. Yurewicz;Gene Oliphant;Jerry L. Hedrick
通过手动解剖将非洲爪蟾卵的三个形态和功能不同的果冻层 J1、J2 和 J3 分离，用二硫苏糖醇溶解，并进行物理化学分析。三层果冻的化学成分各不相同，其中蛋白质为 37% 至 48%，碳水化合物为 63% 至 52%。碳水化合物由己糖胺、半乳糖和岩藻糖组成。每个果冻层中的一些碳水化合物通过 O-糖苷键与蛋白质共价连接，因为在碱存在下观察到碳水化合物部分的β消除。与之前的发现一致，共价连接的硫酸盐位于最内层的果冻层 J1 内。 pH 8.0 下的醋酸纤维素电泳从蛋白质和碳水化合物差异染色的三个果冻层中总共分离出九种大分子成分：J1 产生两种阳极迁移成分；J1 产生两种阳极迁移成分；J1 产生两种阳极迁移成分；J1 产生两种阳极迁移成分；中间层J2产生两种阴极迁移的大分子组分；最外层J3包含五种，三种阳极，两种阴极。十二烷基硫酸钠琼脂糖凝胶电泳分析产生了九个独特的物种，其中六个对蛋白质和碳水化合物进行了一致染色。使用针对总胶冻成分的抗血清进行免疫电泳和 Ouchterlony 双扩散分析，解析了 J1 和 J3 层中一种或两种大分子之间具有交叉反应性的九种不同抗原种类。分析沉降速度离心揭示了八个不同的物种，所有这些物种都表现出超锐利纹图案，并且其 s20,w 值高度依赖于浓度。根据这些分析，非洲爪蟾的卵果冻层由至少 8-9 种不同的大分子物种组成。这些大分子中的大多数与不同的果冻涂层独特地相关。
The three morphologically and functionally distinct jelly coat layers of Xenopus laevis eggs, J1, J2, and J3, were separated by manual dissection, solubilized with dithiothreitol, and physicochemically analyzed. The chemical composition of the three jelly layers varied from 37 to 48% protein and 63 to 52% carbohydrate. The carbohydrate consisted of hexosamines, galactose, and fucose. Some of the carbohydrate in each of the jelly layers was covalently linked to protein through O-glycosidic bonds as beta elimination of the carbohydrate moiety in the presence of alkali was observed. In agreement with a previous finding, covalently attached sulfate was localized within the innermost jelly coat layer, J1. Cellulose acetate electrophoresis at pH 8.0 resolved a total of nine macromolecular components from the three jelly coat layers differentially staining for protein and carbohydrate: J1 yielded two anodically migrating components; the middle layer J2 yielded two cathodically migrating macromolecular components; the outermost layer J3 contained five species, three anodic and two cathodic. Sodium dodecyl sulfate agarose gel electrophoresis analysis yielded nine unique species, six of which stained coincidently for protein and carbohydrate. Immunoelectrophoresis and Ouchterlony double diffusion analyses using antiserum to total jelly components resolved nine different antigenic species with cross-reactivity between one or two macromolecules in layers J1 and J3. Analytical sedimentation velocity centrifugation revealed eight distinct species all of which exhibited hypersharp schlieren patterns and whose s20,w values were highly concentration dependent. On the basis of these analyses, Xenopus laevis egg jelly layers are composed of at least 8-9 distinct macromolecular species. The majority of these macromolecules are uniquely associated with different jelly coat layers.