The role of eukaryotic subtilisin-like endoproteases for the activation of human immunodeficiency virus glycoproteins in natural host cells
The role of eukaryotic subtilisin-like endoproteases for the activation of human immunodeficiency virus glycoproteins in natural host cells
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DOI：
10.1128/jvi.71.2.1036-1045.1997
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发表时间：
1997-02-01
影响因子：
5.4
通讯作者：
Garten, W
中科院分区：
文献类型：
--
作者：
Hallenberger, S;Moulard, M;Garten, W
人类免疫缺陷病毒1型(HIV-1)的前体包膜糖蛋白gp160和HIV-2的gp140的蛋白水解性激活是病毒感染的先决条件，分别导致gp120/gp41和gp125/gp36的形成。卵裂是由细胞内的蛋白水解酶介导的。呋喃是真核枯草杆菌蛋白家族中的一员，已被证明是一种激活HIV的蛋白酶。在这里，我们比较了呋喃和其他哺乳动物枯草杆菌毒素在淋巴细胞和组织中的存在。Northern印迹分析表明，Furin和最近发现的蛋白酶LPC/PC7是仅有的在这些细胞中转录的枯草杆菌蛋白样酶。在不同的淋巴细胞系和单核细胞系中都发现了一种具有酶活性的内切酶。当从痘苗病毒载体中表达时，原蛋白转换酶被正确地加工、运输和分泌到培养基中并具有酶活性。不同的枯草杆菌毒素与HIV包膜前体的共表达表明，Furin和LPC/PC7能够切割HIV-1gp160。此外，两种酶都对HIV-2的包膜前体进行了蛋白水解性处理。Gp140也被PC1在一定程度上切割，然而，它不存在于淋巴细胞中。FURIN和LPC/PC7催化的HIV-1gp160裂解产生具有生物活性的包膜蛋白。总之，在已知的枯草杆菌蛋白家族成员中，只有Furin和LPC/PC7满足在体内激活HIV包膜糖蛋白的蛋白酶的要求。
Proteolytic activation of the precursor envelope glycoproteins gp160 of human immunodeficiency virus type 1 (HIV-1) and gp140 of HIV-2, a prerequisite for viral infection, results in the formation of gp120/gp41 and gp125/gp36, respectively. Cleavage is mediated by cellular proteases. Furin, a member of the eukaryotic subtilisin family, has been shown to be an activating protease for HIV. Here, we compared the presence of furin and other mammalian subtilisins in lymphatic cells and tissues. Northern blot analyses revealed that furin and the recently discovered protease LPC/PC7 were the only subtilisin-like enzymes transcribed in such cells. Furin was identified as an enzymatically active endoprotease present in different lymphocytic, as well as monocytic, cell lines. When expressed from vaccinia virus vectors, the proprotein convertases were correctly processed, transported, and secreted into the media and enzymatically active. Coexpression of different subtilisins with the HIV envelope precursors revealed that furin and LPC/PC7 are able to cleave HIV-1 gp160. Moreover, both enzymes proteolytically processed the envelope precursor of HIV-2. gp140 was also cleaved to some extent by PC1, which is not, however, present in lymphatic cells. Furin- and LPC/PC7-catalyzed cleavage of HIV-1 gp160 resulted in biologically active envelope protein. In conclusion, among the known members of the subtilisin family, only furin and LPC/PC7 fulfill the requirements of a protease responsible for in vivo activation of HIV envelope glycoproteins.