SEQUENCE REQUIREMENTS FOR CLEAVAGE ACTIVATION OF INFLUENZA-VIRUS HEMAGGLUTININ EXPRESSED IN MAMMALIAN-CELLS
SEQUENCE REQUIREMENTS FOR CLEAVAGE ACTIVATION OF INFLUENZA-VIRUS HEMAGGLUTININ EXPRESSED IN MAMMALIAN-CELLS
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DOI：
10.1073/pnas.85.2.324
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发表时间：
1988-01-01
影响因子：
11.1
通讯作者：
WEBSTER, RG
中科院分区：
文献类型：
--
作者：
KAWAOKA, Y;WEBSTER, RG
组织培养系统中血凝素（HA）的裂解与禽流感病毒的毒力有关。为了研究透明质酸切割的结构要求，我们在哺乳动物细胞（CV-1）中表达了一种强毒H5流感病毒的透明质酸基因，并利用位点特异性诱变技术探索了透明质酸的切割位点。在pH值为5时，表达的HA蛋白表现出正常的裂解、转运到细胞膜以及吸附和融合红细胞的能力。HA的位点特异性突变直接证实：(1)该位点的大多数碱性氨基酸对裂解激活至关重要；（ii）除了连接肽序列外，酶识别还需要HA的至少一个其他结构特征；（iii）连接肽的长度可以消除结构特征(s)。
Cleavage of the hemagglutinin (HA) in tissue culture systems has been correlated with virulence of avian influenza viruses. To examine the structural requirements for cleavage of the HA, the HA gene from a virulent H5 influenza virus was expressed in mammalian cells (CV-1), and the cleavage site of the HA was explored by using site-specific mutagenesis. The expressed HA protein exhibited normal cleavage, transport to the cell membrane, and ability to adsorb and to fuse erythrocytes at pH 5. Site-specific mutagenesis of the HA directly established that (i) most of the basic amino acids at this site are critical for cleavage activation; (ii) besides the connecting peptide sequence, at least one other structural feature of the HA is required for enzyme recognition; and (iii) the length of the connecting peptide can abrogate the structural feature(s).