Characterization of the cilia and ciliary membrane proteins of wild-type Paramecium tetraurelia and a pawn mutant.
Characterization of the cilia and ciliary membrane proteins of wild-type Paramecium tetraurelia and a pawn mutant.
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DOI：
10.1083/jcb.89.2.206
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发表时间：
1981-05
期刊：
影响因子：
--
通讯作者：
Gruenstein EI
中科院分区：
文献类型：
--
作者：
Merkel SJ;Kaneshiro ES;Gruenstein EI
从无菌生长的野生型四脲草履虫菌株51s和衍生自该亲本菌株的极端pawn突变菌株d495中分离纤毛和纤毛膜.通过考马斯亮蓝染色检测到60多条分子量为15至大于300 kdalton的蛋白质条带，所述蛋白质条带通过一维SDS聚丙烯酰胺凝胶电泳分离的整个纤毛蛋白质。在考马斯亮蓝染色的膜分离中，约有30条蛋白质条带可见。膜蛋白一维凝胶银染检测到约60条带。野生型和pawn突变株d495膜蛋白的考马斯亮蓝染色分离之间的差异可见于存在于43 kdalton处的条带的量。放射性碘标记的细胞表面蛋白质约15蛋白带在野生型和突变体纤毛。主要的轴丝蛋白未标记。六个膜糖蛋白进行了鉴定，通过染色一维分离与碘化伴刀豆球蛋白A和小扁豆凝集素，两种凝集素，特异性结合葡萄糖和甘露糖残基。通过糖醇乙酸酯衍生物的气相色谱法初步鉴定了纤毛制剂的酸水解产物中存在的两种主要中性糖种类为葡萄糖和甘露糖。
Cilia and ciliary membranes were isolated from axenically grown, wild- type Paramecium tetraurelia strain 51s and from the extreme pawn mutant strain, d495, derived from this parental strain. Over 60 protein bands having molecular weights of 15 to greater than 300 kdaltons were detected by Coomassie Blue staining of whole cilia proteins separated by one-dimensional SDS polyacrylamide gel electrophoresis. About 30 of these protein bands were visible in Coomassie Blue-stained membrane separations. About 60 bands were detected by silver staining of one- dimensional gels of membrane proteins. Differences between Coomassie Blue-stained separations of wild-type and pawn mutant strain d495 membrane proteins were seen in the quantity of a band present at 43 kdaltons. Radioiodination of cell surface proteins labeled approximately 15 protein bands in both wild-type and mutant cilia. The major axonemal proteins were unlabeled. Six membrane glycoproteins were identified by staining one-dimensional separations with iodinated concanavalin A and lentil lectin, two lectins that specifically bind both glucose and mannose residues. Two major neutral sugar species present in an acid hydrolysate of the cilia preparation were tentatively identified as glucose and mannose by gas chromatography of the alditol acetate derivatives.