inhibition of enterovirus 71 entry by peptides targeting i β-sheet of vp1 protein

Download Inhibition of enterovirus 71 entry by peptides targeting I β-sheet of VP1 protein

If you can't read please download the document

Post on 03-Jan-2016

31 views

Category:

Documents

1 download

Embed Size (px)

DESCRIPTION

Inhibition of enterovirus 71 entry by peptides targeting I β-sheet of VP1 protein. Ming-Liang He, Ph.D The Chinese University of Hong Kong. Enterovirus 71 (EV71). A single stranded, non-enveloped RNA virus (~ 7.5 kb in size); EV71 infection causes hand-foot-and-mouth disease (HFMD); - PowerPoint PPT Presentation

TRANSCRIPT

Inhibition of enterovirus 71 entry by peptides targeting I -sheet of VP1 protein

Inhibition of enterovirus 71 entry by peptides targeting I -sheet of VP1 protein

Ming-Liang He, Ph.DThe Chinese University of Hong Kong

Enterovirus 71 (EV71)A single stranded, non-enveloped RNA virus (~ 7.5 kb in size);

EV71 infection causes hand-foot-and-mouth disease (HFMD);

Children under 5 are highly susceptible to EV71 and easily develop central nervous system (CNS) symptoms;

As long as it causes CNS infection or symptoms, leading to neurologic sequelae, including neurological disorders, meningitis, and even death.

Hand, Foot & Mouth Diseasean acute viral infection, commonly seen in infants and children EV71 epidemiology in China

The reported HFMD cases and related death in China from 2007 to 2012.

No drugs, no vaccine available!Lu J et al., Crit. Rev. Microbiol. 2013Principle of Antiviral Block viral entry by peptides, small molecules or antibodies;

Block viral replication;

Block the assembly of virions;

Block the virion secretion;

Disturb the cellular signaling, and therefore block one or more processes in the life cycle of virus.

ACD

The 3D structure of VP1-VP4 complex of EV71 (PDB ID: 3VBS) virion. VP1, VP2, VP3 and VP4 were represented in blue, red, green and yellow, respectively. I- sheet is indicated by a big write arrow.

The VP1 structurecanyona-helix, D- and I- sheet form the canyon, which binds EV71 receptorsD-

The I sheet is highly conserved among EV71 subtypes and suitable for drug design SequenceMWPIAntiviralSP40 (H)QMRRKVELFTYMRFD 2020.49.98StrongSP45 ()AEFTFVACTPTGEVV1935.88.68WeakSP81 ()KSKYPLVVRIYMRMK1912.410.56Very strongSP81-3KSKYPLVVRIYMRMKHVRAWI2675.311.12Very strongTable 1 The physical and chemical parameters of the synthetic peptides

Peptides target I- sheet inhibited cytopathic effects caused by EV71 infections

Cell viability (%)Peptides target I- sheet protected the viability of cells infected by EV71 Folds of inhibiting viral infection (RNA copies, control/peptide)AB**********

Peptides target I- sheet inhibited viral reproduction3CD?2A, 3C?PolyproteinStructureNon-structureIRES51D1B1C1A2A2B2C3A3B3C3DP3VPgIRES guided translation2A3CD?VP0VP3VP1VP4VP2?3C?2A2B2C3A3C3DVPg3BProteinaseHelicasePolymeraseCleavageP1P2The structure of EV71 genome and its encoding proteins EV71 pseudovirus system

AB Folds of inhibiting virus binding (Luc RNA copies of control/peptides)Folds of reducing Luciferase Activities (Control/peptides)******** ********* ***Peptides target I- sheet inhibited viral infectivityPeptides target I- sheet inhibited the binding of virions to host cells (binding at 4C for 30 min; B. Peptides target I- sheet inhibited viral entry as indicatedBy reduced translation expression of luciferase reporter.

SequenceNumberpercentageKSKYPLVVR I YMRMK61161.8%KSKYPLV I R I YMRMK35936.3%KSKYPLL I R I YMRMK40.4%KSKYPLV I RMYMRMK10.1%KSKYPL I VR I YMRMK 20.2%KSKYPL I I R I YMRMK50.5%KSKYPI VVR I YMRMK10.1%KSKYPVVVR IYMRMK10.1%KSKYPLVVRVYMRMK30.3%KSKYPLVVRTYMRMK10.1%KSKYPVV I RIYMRMK10.1%Table 2 The sequences and residue mutation rate of I- in EV71 circulating worldwide The sequences of SP81-3 (same as the strain outbreaked in 2008 at Fuyang city, Anhui Province, China; GenBank: ACS12928.1). A total of 989 VP1 sequences was recorded in GeneBank and EMBL database by February 2013. The mutated residues are shown in red color.The approximately accumulated mutation rate: 247L to 247I/V, 0.3%; 248V to 248I, 0.7%; 37.4%; 251I to 251M/V/T, 0.6%. SequenceAntiviral SP79RTVGSTKSKYPLVVR-SP80GSTKSKYPLVVRIYM+/-SP81 ()KSKYPLVVRIYMRMK+++++SP82YPLVVRIYMRMKHVR++++ SP83VVRIYMRMKHVRAWI++++ SP84VVRIYMRMKHVRAWIPRP-SPC1AAAVVRIYMRMK++SPC2YPLAAAIYMRMK+SPC3YPLVVRAAARMK++SPC4YPLVVRIYMAAA-SPC5YPILIRMYMRMK++++Table 3 Identification of key residues for EV71 infectivityReduction folds of Luciferase Activities (Control/peptides)The antiviral activity of I- mutants

Key residuescanyonArg250, Arg254, Met255 and Lys256 are key residues which locate on theSurface and would involve interaction with EV71 receptorsConclusionThe I -sheet is an important structure to form the convoy of VP1, which involves receptor binding;

The residues in I -sheet are highly conserved among subtypes of EV71, therefore, it is an idea structure for drug target;

Peptide targeting I -sheet potently inhibited EV71 infections;

Residues Arg250, Arg254, Met255 and Lys256 are critical for virion binding to host cells.

Thank you!Chart2368.460.873.26654.53.386.180.685.78796.45.389.288.392.68893.815.693.289.795.974.14.85.5

SPscrSP40SP81SP81-3

Sheet110 uM50 uM100 uM200 uMSPscr33.35.315.66887SP4068.486.189.293.26784.1SP8160.880.688.389.75994.8SP81-373.285.792.695.94.56.43.85.5RNA1050100SPscrSP4045.3126.27 1.72%17.64 2.03%SP8148.9329.35 3.16%18.34 1.55%SP81-33212.84 2.30%6.25 1.04%10501001050100SPscr9796.794.5SPscr33.35.5SP4045.3126.2717.64SP4054.6973.7382.36SP8148.9329.3518.34SP8151.0770.6581.66SP81-33212.846.25SP81-36888.1693.753.311.722.032.683.161.554.52.33.04luc RNA1050100200SPscr959083851050100SP4040381612SPscr51017SP814235249SP40606284SP81-333281410SP81586576SP81-367728633.53.62.94.434.923.233.124.234.442.542.332.563.363.112.43luc1050100200SPscr3412233771226962026935000SP40755423551675794SP81789622581627728SP81-36355153210456871050100SPscr2.508573.5114285735.15428571SP4078.4171493.27152857195.2142857141050100200SP8177.4493.54857142995.351428571SPscr97.491428571496.488571428664.845714285757.9114285714SP81-381.8428495.2285714397.014285714SP4021.58285714296.72857142864.78571428572.2685714286SP8122.566.45142857144.64857142862.08SP81-318.15714285714.37714285712.98571428571.96285714298.27.26.56.34.23.42.53.31.22.31.81.24.33.244.6100 uM4.2SPscr64.852.75SP404.7864.56SP814.6494.764SP81-32.98SPC-123.43SPC-247.712SPC-327.21SPC-460.32456SPC-58.56732100 uMSPscr64.856.5SP81-32.984.3SPC-123.433.24SPC-247.7124.6SPC-327.214.2SPC-460.324562.75SPC-58.567324.56

Sheet16654.58796.48893.874.14.85.5

SPscrSP40SP81SP81-3

Sheet24.53.313.312.682.31.721.723.163.042.032.031.55

SPscrSP40SP81SP81-3uM% of EV71 RNA

Sheet334.434.232.563.54.924.443.363.63.232.543.112.93.122.332.43

SPscrSP40SP81SP81-3uM% of Luciferase RNA

8.24.21.21.27.23.42.32.36.52.51.81.86.33.31.21.2

SPscrSP40SP81SP81-3uM% Relative Luciferase activity

6.52.51.84.33.244.64.22.754.56

100 uM% of relative Luciferase activity

06.504.303.2404.604.202.7504.56

100 uM% of relative luciferase activity

3.312.684.54.51.723.162.32.32.031.553.043.04

SPscrSP40SP81SP81-3uM% of EV71 RNA inhibition

2.5634.434.233.363.54.924.443.113.63.232.54

SPscrSP40SP81SP81-3uM% of relative luciferrase RNA inhibition

1.234.434.22.33.54.923.41.83.63.232.5

SPscrSP40SP81SP81-3uM% of relative luciferase activity inhibition

1050100200SPscr0.97490.96480.648450.5791SP40SP81SP81-3

Recommended

View more >