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STUDY OF BINDING ABILITY OF PHOSPHOROTHIOATE OLIGONUCLEOTIDES WITH PROTEIN

硫代磷酸寡核苷酸与蛋白质结合能力的研究

基本信息

批准号：
08670540
负责人：
SHOJI Yoko
金额：
$ 1.6万
依托单位：
St.Marianna University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1998
项目状态：
已结题

项目摘要

We've recognized that phosphorothioate oligonucleotides (S-ODN) showed potent anti-herpetic activities when target site was limited near the splicing site. It was implied that S-ODN interfered the conservative structure which could be essential for the splicing, thus showed potent anti-herpetic activities. This was one explanation why S-ODN targeted splicing site showed potent anti-herpetic activities. However, it has been reported that G rich sequences reveal sequence non-specific biological activities. S-ODN may reveal another mechanism which might be based on strong protein binding activities. S-ODN strongly bound to serum protein at the ratio of 86.6%, while 21.6% for phosphodiester oligonucleotides (D-ODN). Binding activities which directly bound to the virus was more than 50% for S-ODN, on the other hand it was less than 5% for D-ODN.Thirteen mer S-ODN showed inhibitory effect on virus entry into the cell at the early stage of infection, while D-ODN did not. CD spectrum of S-ODN implied G-quartet structure, however, the relation between biological activities and higher dimension structure could not be clear.Moreover, we tried to enhance the anti-herpetic activities of S-ODN by using cationic liposomes. Cationic liposomes is useful tool to enhance the anti-herpetic activities of D-ODN, while it was not the case for SOD.One of the reason might be strong protein binding activities of S-ODN.From this study, the caution is proposed when S-ODN containing G-rich sequences might involve another mechanism other than antisense manner. We should carefully select the drug delivery system to enhance the biological activities of S-ODN.

硫代磷酸寡核苷酸（S-ODN）在靶位点限制在剪接位点附近时显示出较强的抗疱疹活性。这表明S-ODN干扰了剪切所必需的保守结构，从而表现出较强的抗疱疹活性。这也解释了S-ODN靶向剪接位点的抗疱疹病毒活性。然而，据报道，富含G的序列揭示序列非特异性生物活性。S-ODN可能揭示了另一种机制，该机制可能基于强蛋白结合活性。S-ODN与血清蛋白的结合率为86.6%，而与磷酸二酯寡核苷酸（D-ODN）的结合率为21.6%。S-ODN与病毒的结合活性大于50%，而D-ODN与病毒的结合活性小于5%，S-ODN在感染早期对病毒进入细胞有抑制作用，而D-ODN对病毒进入细胞无抑制作用。S-ODN的CD谱显示其具有G-四联体结构，但其生物活性与高维结构的关系尚不清楚。阳离子脂质体能增强D-ODN的抗疱疹活性，而SOD则不能，其原因之一可能是S-ODN具有较强的蛋白结合活性，因此，当S-ODN含有富含G的序列时，应注意其可能涉及的机制可能不是反义方式。因此，应谨慎选择给药系统，以提高S-ODN的生物活性。