DISSOCIATION OF POLYOMA-VIRUS BY CHELATION OF CALCIUM-IONS FOUND ASSOCIATED WITH PURIFIED VIRIONS
DISSOCIATION OF POLYOMA-VIRUS BY CHELATION OF CALCIUM-IONS FOUND ASSOCIATED WITH PURIFIED VIRIONS
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DOI：
10.1128/jvi.23.3.717-724.1977
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发表时间：
1977-01-01
影响因子：
5.4
通讯作者：
CONSIGLI, RA
中科院分区：
文献类型：
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作者：
BRADY, JN;WINSTON, VD;CONSIGLI, RA
多瘤病毒粒子的X-射线荧光分析表明，Ca 2+与纯化的病毒粒子。用螯合Ca 2+的乙二醇-双-N，N“-四乙酸（EGTA）和还原剂二硫苏糖醇处理纯化的病毒体，引起病毒体解离。电镜观察显示病毒粒子解离至衣壳水平。用150 mM NaCl、10 mM EGTA和3 mM二硫苏糖醇孵育多瘤病毒粒子对于解离反应是最佳的。解离反应的pH范围为7.5-10.5。CsCl密度梯度离心表明EGTA和二硫苏糖醇都是解离发生所必需的;单独使用两种试剂都不能解离病毒。病毒颗粒的主要蛋白质产物在12 S处沉淀。讨论了这些实验与多瘤碱性碳酸盐-碳酸氢盐解离的关系。
Analysis of polyoma virions by X-ray fluorometry demonstrated that Ca2+ was associated with the purified virion. Treatment of purified virions with ethyleneglycol-bis-N,N''-tetraacetic acid (EGTA), which chelates Ca2+ and the reducing agent dithiothreitol, caused the virions to dissociate. EM revealed that the virions were dissociated to the capsomere level. Incubation of polyoma virions with 150 mM NaCl, 10 mM EGTA and 3 mM dithiothreitol was optimum for the dissociation reaction. The pH for the dissociation reaction ranged from 7.5-10.5. CsCl density gradient centrifugation indicated that both EGTA and dithiothreitol were necessary for dissociation to occur; neither reagent alone dissociated the virus. The major protein product fo the dissociated viral particles sedimented at 12S. Relationships between these experiments and the alkaline carbonate-bicarbonate dissociation of polyoma are disscussed.