LEISHMANIA-DONOVANI - IDENTIFICATION OF GLYCOPROTEINS RELEASED BY PROMASTIGOTES DURING GROWTH-INVITRO
LEISHMANIA-DONOVANI - IDENTIFICATION OF GLYCOPROTEINS RELEASED BY PROMASTIGOTES DURING GROWTH-INVITRO
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DOI：
10.1016/0014-4894(88)90067-7
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发表时间：
1988-12-01
影响因子：
2.1
通讯作者：
DWYER, DM
中科院分区：
文献类型：
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作者：
BATES, PA;GOTTLIEB, M;DWYER, DM
代谢标记的杜氏利什曼原虫前鞭毛体的培养物上清液显示含有.apprx。 40 种电泳不同的释放蛋白质化合物。其中.apprx。 20种是含有末端甘露糖残基的糖蛋白，根据它们与伴刀豆球蛋白A-琼脂糖珠的特异性结合来判断。释放的糖蛋白的较小子集与琼脂糖缀合的菜豆、蓖麻和花生凝集素结合。通过伴刀豆球蛋白 A 亲和层析分离释放的含前鞭毛体甘露糖的糖蛋白，并用于免疫兔子。通过溶液中的免疫沉淀和免疫印迹分析，该抗血清识别寄生虫释放的含甘露糖的糖蛋白，包括可溶性酸性磷酸酶。在抗体桥接酶测定中，这种多特异性血清还能够结合溶液中的天然酸性磷酸酶，并在 SDS-PAGE 转印迹上将其桥接至变性酶。尽管该抗血清是针对所有 20 种释放的糖蛋白而产生的，但在琼脂糖凝胶中，其主要沉淀素活性是针对分泌的可溶性酸性磷酸酶。
Culture supernatants of metabolically labeled Leishmania donovani promastigotes were shown to contain .apprx. 40 electrophoretically distinct released protein compounds. Of these .apprx. 20 were glycoproteins which contained terminal mannose residues, as judged by their specfic binding to concanavalin A-agarose beads. Smaller subsets of the released glycoproteins were bound by agarose-conjugated Lens culinaris, Ricinus communis, and peanut lectins. Promastigote mannose-containing released glycoproteins were isolated by concanavalin A affinity chromatography and used to immunize a rabbit. This antiserum recognized the parasite-released mannose-containing glycoproteins, including the soluble acid phosphatase, both by immunoprecipitation from solution and in immunoblot analyses. In a antibody bridged enzyme assay this polyspecific serum was also capable of binding native acid phosphatase out of solution and bridging it to the denatured enzyme on SDS-PAGE transblot. Although this antiserum was raised against all 20 released glycoproteins, in agarose gels its major precipitin activity was against the secreted soluble acid phosphatase.