人博卡病毒2型VP1U突变对sPLA2活性的影响

发布时间：2018-02-26 02:12

本文关键词： 人博卡病毒(HBoV) HBoV VPU 突变体 sPLA活性　出处：《病毒学报》2017年04期 　论文类型：期刊论文

【摘要】：对HBoV2 VP1U蛋白进行原核表达并检测其sPLA_2活性,对其关键氨基酸进行突变,检测突变对sPLA_2活性的影响。构建重组质粒pMD18T-VP1U,选取VP1U蛋白4个关键氨基酸为突变点,分别进行原核表达,蛋白纯化后用sPLA_2试剂盒检测sPLA_2活性,对比突变前后VP1U蛋白sPLA_2活性有无变化。野生型HBoV2VP1U蛋白的sPLA_2活性为0.243μmol/min/mL,具有sPLA_2活性;突变体L19P、P21R、D42H及Y45N蛋白的sPLA_2活性分别为野生型蛋白的1.2%、1.2%、0.82%、0.41%。野生型HBoV2VP1U蛋白具有sPLA_2活性,突变体蛋白均几乎完全丧失了sPLA_2活性,提示19、21、42、45位4个氨基酸是维持sPLA_2活性的关键氨基酸。该研究为靶向抗病毒药物的研究提供了理论基础。
[Abstract]:The HBoV2 VP1U protein was expressed in prokaryotic and its sPLA_2 activity was detected, the key amino acids were mutated and the effect of mutation on sPLA_2 activity was detected. The recombinant plasmid pMD18T-VP1U was constructed and four key amino acids of VP1U protein were selected as mutation sites for prokaryotic expression. After purification, sPLA_2 activity was detected by sPLA_2 kit, and the sPLA_2 activity of VP1U protein was compared before and after mutation. The sPLA_2 activity of wild-type HBoV2VP1U protein was 0.243 渭 mol / min / mL, with sPLA_2 activity. The sPLA_2 activity of the mutant L19PmP21RND42H and Y45N protein were 1.2and 1.2and 1.2and 0.82 of wild-type protein, respectively. The wild type HBoV2VP1U protein had sPLA_2 activity, and the mutant protein almost lost sPLA_2 activity. It is suggested that the 4 amino acids at the 45th position of 19s 21 and 42N are the key amino acids to maintain the activity of sPLA_2, which provides a theoretical basis for the study of targeted antiviral drugs.
【作者单位】：南京医科大学附属儿童医院;
【基金】：江苏省自然科学基金青年基金(项目号:BK2012069),题目:HBOV2VP1独有区氨基酸突变对PLA2活性及细胞因子分泌的影响研究国家自然科学基金青年基金(项目号:81300296),题目:HBoV2VP1U突变对PLA2活性及免疫反应的影响~~
【分类号】：R373

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