| General information | |
| ACovPid: | ACoVP100380 |
| Trivial Name: | P6 |
| Amino Acids Sequence: | GINASVVNIQKEIDRLNEVAKNLNESLIDL |
| Length: | 23 |
| C-Terminal Modification: | None |
| N-Terminal Modification: | None |
| Chemical Modification: | None |
| Peptide Source: | Severe acute respiratory syndrome coronavirus (SARS-CoV) : 694009 |
| Source Description: | From the heptad repeat region 2 of spike protein of SARS-CoV |
| Against Virus: | Severe acute respiratory syndrome coronavirus (SARS-CoV) : 694009 |
| Inhibition Value Type: | IC50 |
| Inhibitory Effect: | 1.04 ± 0.22 |
| Inhibitory Unit: | µM |
| Target Domain Name: | |
| Assay: | Cell–cell fusion assay |
| Assay Description: | The cell fusion inhibition assay is modified from a previously described β-galactosidase reporter gene-based SARS-CoV cell fusion assay (PMID: 14651994, 15492138). VRC(S)8304, a plasmid containing the S gene of SARS-CoV with humanized codon usage, is designated as S(h) in this study (PMID: 15492138). To construct pCDNA3HA-ACE2, total RNA extracted from VeroE6 cells by the Qiagen RNeasy mini kit (Qiagen, Valencia, CA) was subjected to RT using random primers, followed by PCR using the primer pair (NotI-ACE2, 5′-AAGGAAAAAAGCGGCCGCCGATGTCAAGCTCTTCC-3′ and XhoI-ACE2, 5′-CCCGCTC GAGCTAAAAGGAGGTCTGAAC-3′), digestion of the product with NotI and XhoI, and cloning into respective sites of pCDNA3-HA (Invitrogen, Carlsbad, CA). The construct was confirmed by sequencing both junctions of the insert with T7 and SP6 primers. HeLa cells (1 × 10^6) transfected with 2 µg of S(h) (cells #1) by use of lipofectamine 2000 (Invitrogen, Carsblad, CA) were infected with recombinant vaccinia virus expressing β-galactosidase (vCB21R) at a multiplicity of infection (MOI) of 1 at 6 h post-transfection. HeLa cells (1 × 10^6) transfected with 2 µg of pCDNA3HA-ACE2 (cells #2) were infected with recombinant vaccinia virus expressing T7 polymerase (vTF7.3) at a MOI of 1. After adsorption at 37 °C for 2 h and replacement with fresh medium, both cells were incubated at 28 °C overnight, washed with 1× PBS once, trypsinized, washed with 1× PBS twice, and resuspended in DMEM containing 10% FCS. Both cells #1 and #2 (1 × 10^5 cells, each well) were added into 96-well in duplicates, pre-incubated with or without different concentrations of peptides (50 µl per well) at 37 °C for 20 min, and co-cultured at 37 °C for 3 h. Cell lysates were measured for β-galactosidase activity using the Galcto-Start kit (Applied Biosystems, Bedford, MA). The percentage of fusion is the ratio of β-galactosidase activity in the presence of peptide to that in the absence of peptide. |
| Anti-CoV activity in vivo: | |
| Reference: | 18983873 |
| Comment: | |
| 3D structure: | |
| Structure Experiment Verified: | NO |
| Similar Peptides: | ACoVP100242 ACoVP100125 ACoVP100389 ACoVP100119 ACoVP100124 |