General information | |
ACovPid: | ACoVP100361 |
Trivial Name: | 229E-HR2P |
Amino Acids Sequence: | VVEQYNQTILNLTSEISTLENKSAELNYTVQKLQTLIDNINSTLVDLKWL |
Length: | 50 |
C-Terminal Modification: | None |
N-Terminal Modification: | None |
Chemical Modification: | None |
Peptide Source: | Human coronavirus 229E (HCoV-229E) : 11137 |
Source Description: | From the heptad repeat region 2 of spike protein of Human coronavirus 229E |
Against Virus: | Human coronavirus 229E (HCoV-229E) : 11137 |
Inhibition Value Type: | IC50 |
Inhibitory Effect: | 0.31 |
Inhibitory Unit: | µM |
Target Domain Name: | HR2 domain of HCoV-229E |
Assay: | Cell–cell fusion assay |
Assay Description: | It has been reported that HCoV-229E-infected cells could form syncytia with uninfected cells in the presence of trypsin, suggesting, in turn, that the S protein on HCoV-229E-infected cells may mediate the fusion between infected and uninfected cells. Accordingly, to assess the inhibitory activities of 229E-HR1P and 229E-HR2P, we established a cell-cell fusion model. To accomplish this, we used Huh-7 cells, naturally expressing the APN receptor of HCoV-229E on the cell surface, and 293T cells that simultaneously coexpress HCoV-229E S protein on the cell surface and Enhanced Green Fluorescent Protein (EGFP) in the cytoplasm (293T/229E/EGFP) as the target and effector cells, respectively (Figure 3a). After coculture of effector cells (293T/229E/EGFP) and target cells (Huh-7) at 37 °C for 3 to 6 h, we could observe cells that had apparently fused, using fluorescence microscopy, which showed larger size and weaker fluorescence than unfused cells. This can be explained by the diffusion of fluorescence into one or more cells that fuse together (Figure 3b). After staining with 4′,6-diamidino-2-phenylindole (DAPI), we could clearly see several nuclei in one fused cell. However, each of the 293T/229E/EGFP cells cultured alone contained only one single nucleus, suggesting that the effector cells cannot spontaneously fuse with each other or do not undergo propagating divisions to influence the assessment of fusion. Besides, 293T/EGFP cells that express no HCoV-229E S protein on cell surface could not fuse with the target cells, and each of these cells also contained only one nucleus. After 24 h, obvious formation of syncytium occurred, as shown in Figure 3c. Consistent with the previous report, trypsin could effectively promote cell-cell fusion in a time- and dose-dependent manner (Figure 3d). This finding confirms that HCoV-229E S protein can mediate membrane fusion between effector cells and target cells, followed by syncytium formation, suggesting that this cell-cell fusion assay can be used to study HCoV-229E S protein-mediated membrane fusion process and identify HCoV-229E fusion and entry inhibitors. |
Anti-CoV activity in vivo: | The respiratory tract is a key target of HCoV-229E viral infection [14]. Proteolytic enzymes on the surface of respiratory tract mucosa may cause degradation of the peptides [31,32], resulting in attenuation of their antiviral activity. Therefore, the antiviral activity of 229E-HR2P was verified in the respiratory tract of mice intranasally administered with or without 229E-HR2P. One hour later, upper and lower respiratory tract lavage fluids, respectively, were collected to test their inhibitory activities on HCoV-229E S protein-mediated cell-cell fusion. As shown in Figure 7, the upper and lower respiratory tract lavage fluids from mice treated with 229E-HR2P at 1:32 and 1:16 dilutions, respectively, exhibited significant viral membrane fusion-inhibitory activity. These findings suggested that 229E-HR2P may not be degraded by proteolytic enzymes and that the peptide can maintain effective antiviral activity in both upper and lower respiratory tracts against HCoV-229E infection. |
Reference: | 29415501 |
Comment: | |
3D structure: | |
Structure Experiment Verified: | NO |
Similar Peptides: | ACoVP100394   ACoVP100395   ACoVP100396   ACoVP100128   ACoVP100183 |
Target Domain information | |
Target Domain Full Name: | Heptad repeat 2 (HR2) domain of Human coronavirus 229E (HCoV-229E) spike glycoprotein |
Target Type: | glycoprotein |
UniprotID [Sequence]: | P15423 [1031-1127] |
Target Synonyms: | Alternative name(s) for spike glycoprotein: E2 Peplomer protein S glycoprotein |
Target Source: | Human coronavirus 229E (HCoV-229E) : 11137 |
Target Structure: | 5YL9, 5ZHY, 5ZUV, 6ATK, 6U7H, 7CYC, 7CYD |