AMBER TI Tutorials
A simple TIMD tutorial about T4-lysozyme
Introduction to TIMD
This is a TIMD tutorial based on the tutorial written by Thomas Steinbrecher. But some important changes have been made to suit the current AMBER 10 version according to Miranda's tutorial from Simmerling's lab.
Thermodynamic cycle and Method
TI calculations compute the free energy difference between two states A and B by coupling them via a parameters λ that serves as an additional, nonspatial coordinate. This λ formalism allows the free energy difference between the states to be computed as:
Setup of the T4 System
Generation of ligands files
We will use the X-ray structure of T4-L99A from the pdb (after stripping water molecules and unneeded heteroatoms from it: pdb file) as basis to set up our simulation files. We will use two runs of leap to produce four set of parameter and restart files, containing both ligands in the protein bound and solvated states. The first leap run (input file) will produce pdb files of the solvated und neutralized benzene complex and of the benzene ligand in water (complex.pdb and ligand.pdb). From these two additional pdb files are made by renaming the BNZ molecule to PHN and deleting H6 (t4_phn.pdb and phn.pdb). These four pdb files are then used in a second leap run (input file) to generate the *.prm and *.rst files. This yields 4 parameter and 4 rst files
Generate Start Structures
Setup and run MD
After running the simulations on the seawulf cluster, you still have plenty of work to do. The analyze of the data is not so straight and you need to write linux scripts using python or perl to extract the data.