TMPRSS2 and ADAM17 Cleave ACE2 Differentially and Only Proteolysis by TMPRSS2 Augments Entry Driven by the Severe Acute Respiratory Syndrome Coronavirus Spike Protein
TMPRSS2 and ADAM17 Cleave ACE2 Differentially and Only Proteolysis by TMPRSS2 Augments Entry Driven by the Severe Acute Respiratory Syndrome Coronavirus Spike Protein
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DOI：
10.1128/jvi.02202-13
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发表时间：
2014-01-01
影响因子：
5.4
通讯作者：
Poehlmann, Stefan
中科院分区：
文献类型：
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作者：
Heurich, Adeline;Hofmann-Winkler, Heike;Poehlmann, Stefan
Ⅱ型跨膜丝氨酸蛋白酶TMPRSS 2和HAT能切割并激活SARS冠状病毒刺突蛋白（S）进行膜融合。此外，这些蛋白酶切割病毒受体，羧肽酶血管紧张素转换酶2（ACE 2），并提出ACE 2切割增强病毒感染性。然而，没有机制的见解，这一过程中获得的相关性ACE 2切割SARS-CoV S蛋白（SARS-S）激活尚未确定。在这里，我们表明，精氨酸和赖氨酸残基内ACE 2氨基酸697至716是必不可少的切割TMPRSS 2和HAT和ACE 2加工是必需的增强SARS-S驱动的进入这些蛋白酶。相反，ACE 2裂解被排除用于病毒S蛋白的活化。TMPRSS 2的表达增加了可溶性SARS-S的细胞摄取，表明病毒进入的蛋白酶依赖性增强可能是由于病毒体进入靶细胞的摄取增加。最后，发现TMPRSS 2与金属蛋白酶ADAM 17竞争ACE 2加工，但只有TMPRSS 2的切割导致增强的SARS-S驱动进入。总的来说，我们的结果与以前的研究表明，TMPRSS 2和潜在的相关蛋白酶促进SARS-CoV进入两个独立的机制：ACE 2切割，这可能会促进病毒的摄取，和SARS-S切割，激活S蛋白的膜融合。这些观察结果对新疗法的开发具有有趣的意义。此外，他们应该刺激努力，以确定受体裂解是否促进其他冠状病毒的进入，这些病毒使用肽酶作为进入受体。
The type II transmembrane serine proteases TMPRSS2 and HAT can cleave and activate the spike protein (S) of the severe acute respiratory syndrome coronavirus (SARS-CoV) for membrane fusion. In addition, these proteases cleave the viral receptor, the carboxypeptidase angiotensin-converting enzyme 2 (ACE2), and it was proposed that ACE2 cleavage augments viral infectivity. However, no mechanistic insights into this process were obtained and the relevance of ACE2 cleavage for SARS-CoV S protein (SARS-S) activation has not been determined. Here, we show that arginine and lysine residues within ACE2 amino acids 697 to 716 are essential for cleavage by TMPRSS2 and HAT and that ACE2 processing is required for augmentation of SARS-S-driven entry by these proteases. In contrast, ACE2 cleavage was dispensable for activation of the viral S protein. Expression of TMPRSS2 increased cellular uptake of soluble SARS-S, suggesting that protease-dependent augmentation of viral entry might be due to increased uptake of virions into target cells. Finally, TMPRSS2 was found to compete with the metalloprotease ADAM17 for ACE2 processing, but only cleavage by TMPRSS2 resulted in augmented SARS-S-driven entry. Collectively, our results in conjunction with those of previous studies indicate that TMPRSS2 and potentially related proteases promote SARS-CoV entry by two separate mechanisms: ACE2 cleavage, which might promote viral uptake, and SARS-S cleavage, which activates the S protein for membrane fusion. These observations have interesting implications for the development of novel therapeutics. In addition, they should spur efforts to determine whether receptor cleavage promotes entry of other coronaviruses, which use peptidases as entry receptors.