INHIBITION OF PROTEOLYTIC ACTIVATION OF INFLUENZA-VIRUS HEMAGGLUTININ BY SPECIFIC PEPTIDYL CHLOROALKYL KETONES
INHIBITION OF PROTEOLYTIC ACTIVATION OF INFLUENZA-VIRUS HEMAGGLUTININ BY SPECIFIC PEPTIDYL CHLOROALKYL KETONES
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DOI：
10.1016/0042-6822(89)90103-7
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发表时间：
1989-09-01
期刊：
影响因子：
3.7
通讯作者：
KLENK, HD
中科院分区：
文献类型：
--
作者：
GARTEN, W;STIENEKE, A;KLENK, HD
已使用肽基-对硝基苯胺作为显色底物分析培养细胞的裂解物的精氨酸特异性内切蛋白酶。存在于MDBK、MDCK、VERO、BHK和鸡胚细胞中的酶需要赖氨酸-精氨酸或精氨酸-精氨酸对作为切割位点，而绒毛尿囊膜细胞另外含有可在单个精氨酸处切割的活性。研究了肽基氯烷基酮对成对碱性残基特异性蛋白酶激活鸡瘟病毒血凝素的影响。当含有未裂解的血凝素的病毒体与未感染细胞的裂解物一起孵育时，裂解被含有成对碱性残基的肽基氯烷基酮以1 mM的浓度完全抑制。相反，在含有单一精氨酸的化合物中没有抑制活性。当将二元肽基氯烷基酮添加到感染的细胞培养物中时，血凝素的裂解和病毒复制的多个循环在10 mM时被抑制。然而，在完整细胞中的抑制活性增加100- 200倍可以通过N-末端酰化来实现。这些研究表明肽基氯烷基酮作为抗病毒剂的潜在作用。
Lysates of cultured cells have been analyzed for arginine-specific endoproteases using peptidyl-p-nitroanilides as chromogenic substrates. The enzymes present in MDBK, MDCK, VERO, BHK, and chick embryo cells required lysine-arginine or arginine-arginine pairs as cleavage sites, whereas chorioallantoic membrane cells contained, in addition, an activity that could cleave at a single arginine. The effect of peptidyl chloroalkyl ketones on the activation of the fowl plague virus hemagglutinin by the proteases specific for paired basic residues has been investigated. When virions containing uncleaved hemagglutinin were incubated with lysates of uninfected cells, cleavage was completely inhibited by peptidyl chloroalkyl ketones containing paired basic residues at a concentration of 1 mM. In contrast at compound containing a single arginine had no inhibitory activity. When dibasic peptidyl chloroalkyl ketones were added to infected cell cultures, cleavage of hemagglutinin and multiple cycles of virus replication were inhibited at 10 mM. However, a 100- to 200-fold increase of the inhibitory activity in intact cells could be achieved by N-terminal acylation. These studies suggest a potential role of peptidyl chloroalkyl ketones as antiviral agents.