Cloning, characterization, and functional expression of cDNAs encoding glucose transporter proteins from the human parasite Schistosoma mansoni.
Cloning, characterization, and functional expression of cDNAs encoding glucose transporter proteins from the human parasite Schistosoma mansoni.
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DOI：
10.1016/s0021-9258(17)41770-4
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发表时间：
1994-02
期刊：
影响因子：
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通讯作者：
P. Skelly;Jung Woo Kim;J. Cunningham;Charles B. Shoemaker
中科院分区：
文献类型：
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作者：
P. Skelly;Jung Woo Kim;J. Cunningham;Charles B. Shoemaker
曼氏血吸虫的血液寄生成虫，在其穿过皮层运输后，消耗大量的宿主葡萄糖。相关转运蛋白的免疫学或药理学靶向作用可能形成一种靶向控制策略的基础。在这里，我们报告了三种不同的全长cDNA的分离和表征，其预测的蛋白质序列显示出高度的序列和结构相似性，其他动物，植物和细菌的易化扩散转运蛋白。两个cDNA的功能性表达已经通过注射非洲爪蟾卵母细胞与体外衍生的正义链RNA实现。注射的卵母细胞摄取放射性标记的葡萄糖类似物的能力显著高于对照组。S.在卵母细胞中表达的曼氏葡萄糖转运蛋白对D-葡萄糖表现出立体特异性，对不同的己糖具有松弛的特异性，表现出钠非依赖性，并且被根皮素和细胞松弛素B显著抑制。这两个转运蛋白，在卵母细胞中表达，有一个Km为3-O-甲基葡萄糖的1.3和2毫米。第三个葡萄糖转运蛋白同源cDNA似乎来自最近的假基因。这两个功能的S。Mansoni葡萄糖转运蛋白基因在幼虫和成虫雄性和雌性双质体中表达。
The blood-dwelling adult form of the parasitic worm, Schistosoma mansoni, consumes prodigious quantities of host glucose following its transport across the tegument. Immunologic or pharmacologic targeting of the relevant transporter proteins might form the basis of a schistosome control strategy. Here we report the isolation and characterization of three different full-length cDNAs whose predicted protein sequences show a high degree of sequence and structural similarity to the facilitated diffusion transporter proteins of other animals, plants, and bacteria. Functional expression of two cDNAs has been achieved by injection of Xenopus oocytes with in vitro derived sense strand RNA. Injected oocytes have a significantly increased ability to take up radiolabeled glucose analogues over controls. S. mansoni glucose transporters expressed in oocytes exhibit stereospecificity for D-glucose, have relaxed specificity for different hexoses, exhibit sodium independence, and are markedly inhibited by phloretin and cytochalasin B. These two transporters, expressed in oocytes, have a Km for 3-O-methylglucose of 1.3 and 2 mM. A third glucose transporter homologue cDNA appears to derive from a recent pseudogene. Both of the functional S. mansoni glucose transporter genes are expressed in larval and adult male and female schistosomes.