General information | |
ACovPid: | ACoVP100257 |
Trivial Name: | GST‐removed HR2 |
Amino Acids Sequence: | DVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYI |
Length: | 48 |
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: | EC50 |
Inhibitory Effect: | 2.15 |
Inhibitory Unit: | µM |
Target Domain Name: | HR2 domain of SARS-CoV |
Assay: | Pseudotyped virus infection inhibition assay |
Assay Description: | To produce HIV‐luc/SARS pseudotyped virus, 10 µg of HIV‐1 luciferase reporter vector pNL4.3.Luc.E‐R‐luc (HIV‐luc) and 10 µg of codon‐optimized SARS‐CoV S protein expression plasmid (pcTSh) were co‐transfected into 293 T cells by calcium phosphate coprecipitation. The construction of the plasmids have been performeddescribed previously(PMID: 7531918, 15358126). The pseudotyped virus was harvested after 48 h of incubation and purified by ultracentrifugation through a CsCl density gradient at 50,000g for 4 h. The supernatant containing the pseudotyped virus was collected, filtered through a 0.45 µm Millipore‐sized membrane and stored at −80°C until used. To measure luciferase activity, 20‐µl aliquots of the pseudotyped virus were added to 100 µl of Luciferase Assay Reagent (Luciferase Assay Substrate pre‐mixed with Luciferase Assay Buffer, Promega, USA). Luciferase activity was measure 10 s using a Wallac Multilabel 1450 Counter (Perkin‐Elmer, Singapore). GST‐HR2 and His6‐HR1 were mixed in the presence of ADS‐J1, XXT or PBS, followed by immobilization onto glutathione resin. The resin was then washed with PBS and eluted with 10 mM reduced glutathione. The fractions were separated by SDS–PAGE and analyzed by Western blot using anti‐His tag antibodies. When incubated with PBS only, majority of His6‐HR1 was detected in the elution fractions (Fig. (Fig.6A),6A), showing that His6‐HR1 is bound to GST‐HR2. In the presence of ADS‐J1, His6‐HR1 was only found in the flow‐through but not in the elution fractions (Fig. (Fig.6A),6A), whereas XTT had no effect on the interactions and His6‐HR1 was found only in the elution fractions (Fig. (Fig.6B).6B). It indicated that ADS‐J1 interfered with the interactions between His6‐HR1 and GST‐HR2. |
Anti-CoV activity in vivo: | |
Reference: | 18442051 |
Comment: | |
3D structure: | |
Structure Experiment Verified: | NO |
Similar Peptides: | ACoVP100125   ACoVP100119   ACoVP100118   ACoVP100121   ACoVP100120 |
Target Domain information | |
Target Domain Full Name: | Heptad repeat 2 (HR2) domain of Severe acute respiratory syndrome coronavirus (SARS-CoV) spike glycoprotein |
Target Type: | glycoprotein |
UniprotID [Sequence]: | P59594 [1145-1184] |
Target Synonyms: | Alternative name(s) for spike glycoprotein: E2 Peplomer protein S glycoprotein |
Target Source: | Severe acute respiratory syndrome coronavirus (SARS-CoV) : 694009 |
Target Structure: | 1WNC, 1WYY, 1ZV7, 1ZV8, 1ZVB, 2AJF, 2BEQ, 2BEZ, 2DD8, 2FXP, 2GHV, 2GHW, 2RUM, 2RUN, 2RUO, 3BGF, 3D0G, 3D0H, 3D0I, 3SCI, 3SCJ, 3SCK, 3SCL, 5WRG, 5X4S, 5X58, 5X5B, 5XJK, 5XLR, 5ZVM, 6ACC, 6ACD, 6ACG, 6ACJ, 6ACK, 6CRV, 6CRW, 6CRX, 6CRZ, 6CS0, 6CS1, 6CS2, 6M3W, 6NB6, 6NB7, 6VW1, 6WAQ, 7JN5 |