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1. 重庆医科大学附属儿童医院P2实验室
2. 重庆医科大学附属儿童医院P2实验室 重庆400014 江西省儿童医院
纸质出版日期:2007,
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[1]黄路,赵晓东,张琴,赵耀,丁媛.人偏肺病毒分离株蛋白质特性的初步探讨[J].重庆医科大学学报,2007(10):1016-1019.
HUANG Lu, et al. Preliminary study on proteins of human metapneumovirus chinese isolate[J]. Journal of Chongqing Medical University, 2007,(10):1016-1019.
目的:初步明确人偏肺病毒(Human metapneumovirus
hMPV)主要蛋白质的基本特性。方法:以纯化的hMPV重庆分离株CHN05-01灭活病毒为抗原
免疫家兔获得抗血清。用此抗血清为一抗通过免疫印迹法检测hMPV蛋白。用NetNGlyc 1.0 Server、NetOGlyc 3.1 Server、NetPhos 2.0 Server分析hMPVA型标准株CAN97-83氨基酸序列
预测hMPV蛋白氨基酸序列中可能的糖基化、磷酸化位点。结果:ELISA检测显示所制备的抗血清与hMPV的蛋白反应效价达1:500以上
可有效识别hMPV蛋白。软件分析示G蛋白糖基化位点最多
而P蛋白磷酸化位点最多。以兔抗hMPV血清为一抗的免疫印迹法提示CHN05-01的G蛋白糖基化水平较一致
与CAN99-80和CAN99-81不同
其分子量介于55至72ku之间
约68ku;F蛋白F1亚单位分子量介于40至55ku
约为48ku。结论:初步明确了hMPV两个主要膜表面糖蛋白的分子量和糖化特点
为后续蛋白功能研究奠定了基础。
Objective:Human metapneumovirus (hMPV)
initially described in 2001
is an enveloped RNA virus of the genus Metapneumovirus
subfamily Pneumovirinae
family Paramyxoviridae. Here we sought to clarify basic features of human metapneumovirus proteins. Methods:Rabbits were immunized with inactivated virons of hMPV Chinese first isolate
CHN05-01
to yield anti-hMPV antiserum. Antiserum was used as primary antibody to detect hMPV proteins by Western blotting. NetNglyc1.0 server
NetOglyc 3.1server and the NetPhos 2.0 server were applied for predicting potential glycosylation and phosphorylation sites of proteins of prototype virus of type A
CAN97-83. Results:The highest reactive titer of the antiserum with hMPV antigens reached 1:500 in ELISA. Potential glycosylation sites of G protein and phosphorylation sites of P protein were greatest among all hMPV proteins. G protein was shown a narrow band with molecular weight between 55 and 72kDa (approximately 68kDa)
indicating its glycosylation level being consistent and remarkably different from that of CAN99-80 and CAN99-81. F1 subunit of fusion protein displayed molecular weight between 40 and 55kDa (approximately 48 kDa)
which is consistent with previous reports. Conclusion:Basic features of two major membrane proteins of Chinese human metapneumovirus isolate were clarified
which will benefit future studies on protein funtion and pathogenesis of this virus.
偏肺病毒F蛋白G蛋白质抗血清
MetapneumovirusFusion proteinAttachment proteinAntiserum
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