3Coronary artery diseases and CETP Atherosclerosis - leading cause of mortality in industrialized nations.Recent studies have shown that Cholesteryl Ester Transfer Protein (CETP), which modulates the neutral lipid profile in the body between HDL and LDL, is an effective target.Koivuniemi, A., T. Vuorela, et al. (2012). PLoS Computational Biology 8(1).
4Cholesteryl ester transfer protein Boomerang shaped protein molecule with a hydrophobic tunnel.Transfers lipids between High Density Lipoprotein (HDL) and Low Density Lipoprotein (LDL)Inhibiting CETP caused reduction of plaques in the arterial walls of rabbits.Qiu, X., A. Mistry, et al. (2007) Nature structural & molecular biology 14(2):Zhang, L., F. Yan, et al. (2012) Nature chemical biology 8(4):
5Currently available inhibitors of CETP ObjectiveTo study the mechanism of CETP inhibition by the3,5-bis(trifluoromethy)phenyl derivatives
7Molecular dynamics Simulations Biomolecular simulations – Computational microscope for molecular biology.Molecular Dynamics is based on Newton’s laws of motion-𝜵E=FiNon-bondedBondedComputer Simulations of Liquids Allen and Tilldeseley
8Why do we need HPCE for MD? System under study:Protein of 7500 atoms protein522 lipid atoms81 atoms of inhibitor (variable)268 Na+ 279 Cl- ionsatoms from waterTotal of over atoms
9Software used: NAMD2.9System scales up to 11ns/day with 256 CPUs in VIRGO
10Flow Chart Ligand structure optimization by Gaussian 3.0 Docking of the inhibitors to CETP by AutoDock4Molecular Dynamics Simulations by NAMD2.9 with CHARMM36 forcefieldVisualization and Analysis using VMD and GROMACS
11ResultsRMSD as a function of time reveals that the systems have stabilized.RMSF results show that the N-terminal distal region of the inhibitor bound CETP has higher atomic fluctuations in comparison to the substrate-bound complex.
12Dynamic Cross Correlation Analysis Anti-correlationPositive -correlationSubstrate bound CETPTorcetrapib bound CETPAnacetrapib bound CETPEvacetrapib bound CETPInhibitor binding increases the anti-correlation between the binding site and distal regions of CETPGrant, Rodrigues, ElSawy, McCammon, Caves, (2006) Bioinformatics 22,
13Principal Component Analysis Inhibitor bound complexes are more dynamic than the substrate bound complexThe above plot reveals that the first 20 Eigen vectors are required to describe 80% of the overall dynamics of the protein.Pronk, S., S. Páll, et al. (2013). Bioinformatics (Oxford, England) 29(7):
16PCA : Porcupine plotsPC1PC2Substrate boundInhibitor boundInhibitor bound CETP shows greater extent of twisting in PC2Humphrey, W., Dalke, A. and Schulten, K., "VMD - Visual Molecular Dynamics", J. Molec. Graphics, 1996, vol. 14, pp
17APBS Analysis Substrate-bound 0 ns 50 ns 100 ns 150 ns The N-terminal distal region of both the complexes become predominantly electropositive over time .The inhibitor bound complex becomes relatively more electropositive.Torcetrapib-boundAnacetrapib-boundEvacetrapib-boundBaker NA, Sept D, Joseph S, Holst MJ, McCammon JA. Electrostatics of nanosystems: application to microtubules and the ribosome. Proc. Natl. Acad. Sci. USA 98,
18In conclusion..Inhibitor binding affects the atomic fluctuations at the distal regions of CETP.Inhibitor binding increases the anti-correlation between the binding site and distal regions of CETP.PCA reveals increased dynamics of the inhibitor bound complexes.Dynamic Cross Correlation Analysis and PCA are indicative of differential twists in the inhibitor-bound systems.This twisting results in the progressive exposure of electropositive residues in the N-terminal distal region.Improved electro-positivity at the HDL –sensing N-terminus supports the existing hypothesis of high affinity of inhibitor bound complexes towards HDL having electronegative surface thereby resulting in the formation of non-productive CETP-HDL complex.**Clark RW, Ruggeri RB, Cunningham D, Bamberger MJ J Lipid Res 2006;47:537–552
19Acknowledgements THANK YOU… Principal Investigator : Dr.Sanjib SenapatiMembers of the Computational Biophysics Lab, IITMHPCE Team:Mr. V. RavichandranMrs.P.GayathriTHANK YOU…