Surface-specific iodination of membrane proteins of viruses and eucaryotic cells using 1,3,4,6-tetrachloro-3alpha,6alpha-diphenylglycoluril.
Surface-specific iodination of membrane proteins of viruses and eucaryotic cells using 1,3,4,6-tetrachloro-3alpha,6alpha-diphenylglycoluril.
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使用 1,3,4,6-四氯-3α,6α-二苯基甘脲对病毒和真核细胞的膜蛋白进行表面特异性碘化。
DOI：
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发表时间：
1978
期刊：
影响因子：
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通讯作者：
Fox Cf
中科院分区：
文献类型：
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作者：
M. Ma;Fox Cf
用碘试剂1，3，4，6-tetrachloro-3alpha，6alpha-diphenylglycouril(氯甘脲)选择性地标记膜表面蛋白，在以下系统中进行了研究：包膜病毒(仙台病毒和新城疫病毒)、人红细胞和在悬液(EL-4)和单层培养(BHK-21)中增殖的有核细胞。描述了在保持完全的病毒完整性或细胞活力的同时，对表面蛋白进行特别碘化的条件。氯甘脲标记表面膜蛋白的方法与乳过氧化物酶和氯胺-T法的比较表明，氯甘脲方法有许多优点：它常规地产生3-17倍的比放射性而不牺牲病毒或细胞的完整性，它在技术上使用更简单，它不需要添加外来的蛋白质来启动反应，也不需要强大的还原剂来终止反应。氯甘脲在病毒和细胞蛋白的非载体标记中也被证明是氯胺-T的有效替代品。膜蛋白样品在碘化前用洗涤剂十二烷基硫酸钠溶解，或在高碘浓度下标记，而不需要事先增溶。使用氯甘脲产生的最终比放射性等于或大于氯胺-T法产生的比放射性。氯甘脲的有效性、使用简单性和通用性使其成为膜系统表面特异性和非载体标记的碘化试剂。
: The use of the iodinating reagent 1,3,4,6-tetrachloro-3alpha,6alpha-diphenylglycouril (chloroglycoluril) to selectively label membrane surface proteins was investigated with the following systems: enveloped viruses (Sendai and Newcastle disease viruses), human erythrocytes, and nucleated cells propagated both in suspension (EL-4) and in monolayer culture (BHK-21). Conditions are described for specifically iodinating surface proteins while maintaining full virus integrity or cell viability. Comparison of the chloroglycoluril method with the lactoperoxidase and chloramine-T methods for labeling surface membrane proteins shows that the chloroglycoluril method has a number of advantages: It routinely produces a 3- to 17-fold greater specific radioactivity without sacrificing viral or cellular integrity, it is technically simpler to use, it does not require the addition of extraneous protein to initiate the reaction nor a strong reducing reagent to terminate it. Chloroglycoluril also proved to be an effective substitute for chloramine-T in the nonvectorial labeling of viral and cellular proteins. Membrane protein samples were solubilized with the detergent sodium dodecyl sulfate before iodination or labeled in the presence of high iodide concentrations without prior solubilization. The resulting specific radioactivities generated by the use of chloroglycoluril were equal to or greater than those generated by the chloramine-T method. The effectiveness, simplicity of use, and versatility of chloroglycoluril recommend it as an iodinating reagent for both surface-specific and nonvectorial labeling of membrane systems.