人博卡病毒2型VP1U突变对sPLA2活性的影响  

Influence of HBoV 2 VP1U Mutations on sPLA_2 Activity

作　　者：庄著伦[1] 杨静[1] 闫坤龙[1] 程卫霞[1] 

机构地区：[1]南京医科大学附属儿童医院,南京210008

出　　处：《病毒学报》2017年第4期501-504,共4页Chinese Journal of Virology

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~~

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。HBoV2VP1Uproteins and its four mutants were expressed in a prokaryotic system,and their secreted PLA2（sPLA_2）activities investigated.The recombinant plasmid pMD18T-VP1 U was constructed,and four key amino acids in the protein were selected for mutation points.These five proteins were expressed and purified,and then their sPLA_2 activity detected using a commercial kit.Compared with bee venom sPLA_2-positive controls the sPLA_2 activity of wild-type HBoV2 VP1 U protein was 0.243μmol/min/mL,which suggested that HBoV2 VP1 U protein had sPLA_2 activity.The mutants L19 P,P21R,D42 Hand Y45Nhad 1.2%,1.2%,0.82% and 0.41% of the sPLA_2 activity of wild-type HBoV2VP1 U protein,respectively.These results suggested that the VP1 Uprotein had sPLA_2 activity,and mutation of one of the amino acids（L19P,P21 R,D42Hand Y45N）in VP1 Ualmost eliminated the sPLA_2 activity of HBoV2VP1 U.These data indicated that these residues are crucial for the sPLA_2-like activity of HBoV2VP1 U.VP1Ucould be a target for the development of anti-viral drugs against HBoV2.

关 键 词：人博卡病毒(HBoV) HBoV2 VP1U 突变体 sPLA2活性 

分 类 号：R373.2[医药卫生—病原生物学]