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“Interação entre peptídeos virais de fusão com membranas modelo”. Danilo Olivier – FFCLRP/USP

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“Interação entre peptídeos virais de fusão com membranas modelo”, presented at http://3whpc.lcca.usp.br

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“Interação entre peptídeos virais de fusão com membranas modelo”. Danilo Olivier – FFCLRP/USP

  1. 1. Interaction of Dengue Fusion Peptides with model membrane systems Danilo Olivier Amando Ito April 16, 2015 Danilo Olivier Amando Ito Dengue Peptides in interaction with model membrane
  2. 2. Dengue Fever and Dengue Virus Aedes aegypti Size: 50nm Flaviviruses, Yellow Fever 4 Different Serotypes 1 1 Source: The Journal of General Virology, 87(Pt 10), 2755–66, (2006). Danilo Olivier Amando Ito Dengue Peptides in interaction with model membrane
  3. 3. Infection and Fusion Process Figure : Mechanism of fusion.2 Endocytosis Virus mature in low pH Fusion Peptide is exposed 2 Source: The Journal of General Virology, 87(Pt 10), 2755–66, (2006). Danilo Olivier Amando Ito Dengue Peptides in interaction with model membrane
  4. 4. Viral Fusion Peptide and Membrane Models Peptides: Putative Fusion Peptide: DEN II.98-112 Large Sequence: DEN II.88-123 KRFVC KHSMV DRGWG NGCGL FGKGG IVTCA MFTCKK Figure : Top: Dengue Peptide in NewCartoon and Tryptophan in CPK representation. Bottom: Dengue Virus Peptide sequence. Charged polar (green), Hydrophobic (blue) and non-charged polar (red). PDB Code:1OK8. Models: Large Unilamellar Vesicles (Experiments) - Lipid Bilayers (In Silico Experiments) Lipids: DMPC, DMPG Danilo Olivier Amando Ito Dengue Peptides in interaction with model membrane
  5. 5. MD: DEN II.88-123 in interaction with DMPC bilayer Danilo Olivier Amando Ito Dengue Peptides in interaction with model membrane
  6. 6. MD: Equilibrium of DMPC bilayer Figure : RMSD of the system during the simulation. Danilo Olivier Amando Ito Dengue Peptides in interaction with model membrane
  7. 7. MD: Root Mean Square Fluctuation 90 100 110 120 Residue Number 0 5 10 15 RMSF(nm) DEN.II 88-123 DEN.II 88-123 + DMPC Figure : Root mean square fluctuation in solution and DMPC. Danilo Olivier Amando Ito Dengue Peptides in interaction with model membrane
  8. 8. MD: Electronic Density Profile 0 0,1 0,2 0,3 0,4 0,5 Density(e - /Å 3 ) Total DMPC Water Peptide Trp -40 -20 0 20 40 z-Axis ( Å ) 0 0,0005 0,001 Danilo Olivier Amando Ito Dengue Peptides in interaction with model membrane
  9. 9. MD: Order Parameter 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Carbon Atom Number ( # ) 0 0,05 0,1 0,15 0,2 0,25 |SCD | DMPC DMPC + Pep Figure : Order Parameter after 180ns of simulation. Danilo Olivier Amando Ito Dengue Peptides in interaction with model membrane
  10. 10. MD: Area per lipid 0 20 40 60 80 100 120 140 160 180 Time (ns) 54 56 58 60 62 64 66 68 Area(Å 2 ) Figure : Area per lipid during the simulation. Danilo Olivier Amando Ito Dengue Peptides in interaction with model membrane
  11. 11. MD/FLU: Anisotropy and Rotational Correlation A = A1 ∗ exp(−x/θ1) + A2 ∗ exp(−x/θ2) + A∞ (1) Table : Comparison between exponential fit for fluorecence anisotropy and rotational correlation for Trp. System Fit A1 θ1(ns) A2 θ2(ns) A∞ DEN II.88-123 Exp. 0,11 0,35 0,02 2,03 -0,018 Comp. 0,12 0,35 0,15 3,70 -0,001 DEN II.88-123 + DMPC Exp. 0,21 0,04 0,11 0,33 0,018 Comp. 0,15 0,59 0,17 3,06 -0,001 Danilo Olivier Amando Ito Dengue Peptides in interaction with model membrane
  12. 12. MD/FLU: Anisotropy and Rotational Correlation A = A1 ∗ exp(−x/θ1) + A2 ∗ exp(−x/θ2) + A∞ (1) Table : Comparison between exponential fit for fluorecence anisotropy and rotational correlation for Trp. System Fit A1 θ1(ns) A2 θ2(ns) A∞ DEN II.88-123 Exp. 0,11 0,35 0,02 2,03 -0,018 Comp. 0,12 0,35 0,15 3,70 -0,001 DEN II.88-123 + DMPC Exp. 0,21 0,04 0,11 0,33 0,018 Comp. 0,15 0,59 0,17 3,06 -0,001 Table : Steady state anisotropy. 88 - 123 ABuffer 0.021 ADMPC 0.034 Danilo Olivier Amando Ito Dengue Peptides in interaction with model membrane
  13. 13. MD/FLU: Secondary Structure of DEN II.88-123 during interaction with DMPC bilayer. Danilo Olivier Amando Ito Dengue Peptides in interaction with model membrane
  14. 14. MD/FLU: Secondary Structure of DEN II.88-123 during interaction with DMPC bilayer. Figure : Secondary Structure colors: Yellow: Coil, Orange: Bridge, Purple: Extented, Green: Turn, Blue: 3-10 Helix. Danilo Olivier Amando Ito Dengue Peptides in interaction with model membrane
  15. 15. Final Configuration Danilo Olivier Amando Ito Dengue Peptides in interaction with model membrane
  16. 16. Preliminary Conclusions Peptide interacts with lipid bilayer; No significant changes in the lipid bilayer; Differences in fluorecence anysotropy decay of Trp; Low agreement between experiment - simulation for the anisotropy in DMPC; DEN II.88-123 in membrane is hindered compare with peptide in solution; Danilo Olivier Amando Ito Dengue Peptides in interaction with model membrane
  17. 17. Acknowledgement USP - Universidade de São Paulo Rice University CNPq Danilo Olivier Amando Ito Dengue Peptides in interaction with model membrane

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