An amphipathic sulphated glycoconjugate of Leishmania: characterization with monoclonal antibodies.
An amphipathic sulphated glycoconjugate of Leishmania: characterization with monoclonal antibodies.
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利什曼原虫的两亲性硫酸化糖缀合物：用单克隆抗体进行表征。
DOI：
10.1002/j.1460-2075.1984.tb02130.x
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发表时间：
1984
期刊：
影响因子：
--
通讯作者：
Goding,JW
中科院分区：
文献类型：
--
作者：
Handman,E;Greenblatt,CL;Goding,JW
由两种单克隆抗体鉴定的热带利什曼原虫的主要糖缀合物被证明是一种外向的两亲性膜抗原，脱落到寄生虫生长的培养基中。该分子可以用[3H]葡萄糖、[3H]半乳糖、[32P]磷酸盐和[35S]硫酸盐进行代谢标记。它在 SDS-聚丙烯酰胺凝胶中电泳时作为多分散带迁移，跨越对应于表观摩尔的凝胶区域。重量。 20 000‐67 000 道尔顿。使用半乳糖氧化酶和[3H]-硼氢化钠可以在活体前鞭毛体的表面上标记一个明显相同的分子家族。该分子在几个小时内被释放到上清液中。检测 3H 或 35S 标记分子需要放射自显影曝光数天，但使用单克隆抗体包被的硝酸纤维素的新型印迹技术可以在电荷转移电泳、蛋白质印迹和斑点印迹中快速检测该分子。糖缀合物在琼脂糖中的电泳迁移率相对于其在 Triton X-100 中的迁移率在脱氧胆酸盐存在下增加，而在十六烷基三甲基溴化铵存在下降低，表明与插入膜的脂质双层一致的两亲特性。使用斑点印迹技术，在 LRC-L137 的所有毒力和无毒克隆以及热带乳杆菌的另外两个分离株（LRC-L287 和 LRC-L251）中检测到糖复合物，但在杜诺瓦尼乳杆菌或墨西哥乳杆菌中未检测到，这与之前描述的抗体特异性一致。然而，本文使用的一般方法表明，L. donovani (LRC-L52) 和 L. mexicana (LRC-L94) 合成了相似但抗原性不同的糖复合物。
A major glycoconjugate of Leishmania tropica major identified by two monoclonal antibodies was shown to be an externally oriented, amphipathic membrane antigen shed into the culture medium in which the parasites grow. This molecule could be labelled metabolically with [3H]glucose, [3H]galactose, [32P]phosphate and [35S]sulphate. It migrated as a polydisperse band upon electrophoresis in SDS‐polyacrylamide gels, spanning the region of the gel corresponding to an apparent mol. wt. of 20 000‐67 000 daltons. An apparently identical family of molecules could be labelled on the surface of living promastigotes using galactose oxidase and [3H]‐sodium borohydride. This molecule was shown to be released into the supernatant over a period of several hours. Detection of the 3H‐ or 35S‐labelled molecule required several days exposure of autoradiographs, but a novel blotting technique using nitrocellulose coated with monoclonal antibody allowed rapid detection of the molecule in charge shift electrophoresis, Western blotting and dot blotting. The electrophoretic mobility of the glycoconjugate in agarose relative to its mobility in Triton X‐100 was increased in the presence of deoxycholate, and decreased in the presence of cetyl trimethyl‐ammonium bromide, indicating amphipathic properties consistent with insertion into the lipid bilayer of the membrane. Using the dot‐blotting technique the glycoconjugate was detected in all virulent and avirulent clones of LRC‐L137 and in two additional isolates of L. tropica major (LRC‐L287 and LRC‐L251), but not in L. donovani or L. mexicana, consistent with the previously described specificity of the antibodies. However, the general approaches used in this paper showed that L. donovani (LRC‐L52) and L. mexicana (LRC‐L94) synthesize a similar, but antigenically distinct glycoconjugate.
糖蛋白和蛋白聚糖的生物化学
影响因子：
--
作者：
W. Lennarz
通讯作者：
W. Lennarz
联合国开发计划署/世界银行/世界卫生组织热带病研究和培训特别规划。
DOI：
--
发表时间：
1978
期刊：
Papua and New Guinea medical journal
影响因子：
--
作者：
A. Lucas
通讯作者：
A. Lucas
复杂碳水化合物：它们的化学、生物合成和功能
影响因子：
3.9
作者：
R. Hughes
通讯作者：
R. Hughes