Latest revision as of 22:22, 5 May 2024

Introduction

DOCK6 is a great tool that helps us understand ligand binding poses and relative binding energies. When DOCK6 is combined with another computational method known as molecular dynamics, we can calculate free energies of binding and probe how the ligand behaves in the binding pocket. In this tutorial we will walk through the use of AMBER16 to perform and analyze molecular dynamic simulations of PDBID 1NDV.

Getting Started

Before starting simulations it is important to create our directories so that we can keep our simulations organized. Create the following directories:

000.paramters
001.tleap_build
002.equilbration
003.production
004.analysis
zzz.master

We will use the zzz.master directory to store our initial mol2 files and other files we may use that do not belong in another directory. Once you have your directories created you can begin to prepare they system for simulation.

Before Simulation

Structure

We will need mol2 and pdb files of the ligand and receptor separately for these simulations. If you followed the 1NDV DOCK6 VS tutorial you will already have these files and can move them to the zzz.master directory. If you did not follow that tutorial the steps will be listed below, but please refer to it if you require a more in depth tutorial. Please note that the receptor file should be without charges and hydrogens, while the ligand will need the charges and hydrogens.

We will start by generating our receptor file. Open the PDB of 1NDV in Chimera and follow these steps.

1) Select an atom on the receptor and use the entire up arrow until the entire protein is selected
2) Select -> Invert (all models)
3) Actions -> Atoms/Bonds -> delete

This should leave us with just the receptor atoms. Save this as a mol2 file and a pdb file as "1ndv_protein_noH_noCharge" DO NOT add charges or hydrogens.

We will now create the ligand file in a similar fashion.

1) Select an atom on the ligand and use the entire up arrow until the entire protein is selected
2) Select -> Invert (all models)
3) Actions -> Atoms/Bonds -> delete

Before we use this file we must add hydrogens and charges.

To add Hydrogens: Tools -> Structure Editing -> Add Hydrogens
To add Charges: Tools -> Structure Editing -> Add Charges (for this ligand we will use an overall charge of 0)

Be sure to reference the PDB file to ensure that Chimera adds hydrogens in the correct places. Once charges and hydrogens are added you can save the mol2 and pdb files as "1ndv_ligand_addH_addCharge" Place both of these files in the zzz.master directory.

Force Field Parameters

Before we can use TLEap, we need to generate forcefield parameters for our ligand. We will use the antechamber program to do this. Move to the 000.paramters directory and use the following command to generate the forcefield parameter file.

antechamber -i ../zzz.master/1ndv_ligand_addH_addCharge.mol2 -fi mol2 -o 1ndv_ligand_antechamber.mol2 -fo mol2 -at gaff -c bcc -rn LIG

When running this you may encounter some warnings such as:

Warning: The number of bonds (3) for atom (ID: 1, Name: N1) does not match the connectivity (2) for atom type (N.ar) defined in CORR_NAME_TYPE.DAT.

Be sure to double check your ligand has the correct bond connectivity, and then these warnings may be safely ignored. You may also encounter some errors that prevent antechamber from running. It may require you to enter the mol2 file and edit the atom types to resolve them. Once antechamber has successfully run, run the following command to make our modified forcefield parameters:

parmchk2 -i 1ndv_ligand_antechamber.mol2 -f mol2 -o 1ndv_ligand.am1bcc.frcmod

TLEap

Now that we have our modified forcefield parameters we can use TLEap to solvate and prepare the system for simulation. Note that this system has no disulfide bonds, but an example of where to place these in the TLEaP file are included. Create a file called "tleap.build.in" and add the folowing commands:

#!/usr/bin/sh 
###load protein force field
source leaprc.protein.ff14SB
###load GAFF force field (for our ligand)
source leaprc.gaff
###load TIP3P (water) force field
source leaprc.water.tip3p
###load ions frcmod for the tip3p model
loadamberparams frcmod.ionsjc_tip3p 
###needed so we can use igb=8 model
set default PBradii mbondi3

###load protein pdb file
rec=loadpdb ../001.structure/1ndv_built.pdb
 
#THIS IS WHERE YOU WOULD DEFINE DISULFIDE BONDS
#NUMBERING SHOULD MATCH INPUT PDB FILE
#bond rec.Res#.SG rec.Res#.SG
###load ligand frcmod/mol2
loadamberparams ../002.parameters/1ndv_ligand.am1bcc.frcmod
lig=loadmol2 ../002.parameters/1ndv_ligand_antechamber.mol2
 
###create gase-phase complex
gascomplex= combine {rec lig}
 
###write gas-phase pdb
savepdb gascomplex 1ndv.gas.complex.pdb

###write gase-phase toplogy and coord files for MMGBSA calc
saveamberparm gascomplex 1ndv.complex.parm7 1ndv.gas.complex.rst7
saveamberparm rec 1ndv.gas.receptor.parm7 1ndv.gas.receptor.rst7
saveamberparm lig 1ndv.gas.ligand.parm7 1ndv.gas.ligand.rst7
 
###create solvated complex (albeit redundant)
solvcomplex= combine {rec lig}
 
###solvate the system
solvateoct solvcomplex TIP3PBOX 12.0
 
###Neutralize system
addions solvcomplex Cl- 0
addions solvcomplex Na+ 0
 
#write solvated pdb file
savepdb solvcomplex 1ndv.wet.complex.pdb
 
###check the system
charge solvcomplex
check solvcomplex
 
###write solvated toplogy and coordinate file
saveamberparm solvcomplex 1ndv.wet.complex.prmtop 1ndv.wet.complex.rst7
quit

Be sure that all files are generated, especially the wet.complex files, and review the tleap_output.txt file to ensure that no errors were encountered. We will also load the wet.complx files into Chimera to make sure TLEaP ran properly. Download the 1ndv.wet.complex.prmtop and 1ndv.wet.complex.rst7. Now in Chiemra:

Tools → MD/Ensemble Analysis → MD Movie

We will use the rst7 file as our trajectory for this step. It should load an image similar to this.

Now lets make sure that the rest of the waters were added properly and that our complex is in the solvent box. To do this:

1) Select → Chain → water
2) Actions → Atoms/Bond → Show

This should show all the waters added by TLEaP and should look similar to this.

Minimization and Equilibration

Before we run a production simulation, we must first minimize and equilibrate the system. We will do this with a 9-step procedure. This process will minimize the energy of the structure, slowly heat the system to the desired temperature, and then equilibrate the system at that temperature. First navigate the the 002.equiilibration directory. As we create the input files pay attention to the "restrainmask" variable. We want to restrain the protein and ligand atoms but not the hydrogens added by TLEaP so this will be set to ":1-350 & !@H=" for our system but be sure to change this is you are using a different receptor and ligand. This will change to just the protein atoms i.e. ":1-349 & !@H=" after step 7.

Create a file 01.min.mdin and add the following lines:

Minmize all the hydrogens
 &cntrl
 imin=1,           ! Minimize the initial structure
 maxcyc=5000,    ! Maximum number of cycles for minimization
 ntb=1,            ! Constant volume
 ntp=0,            ! No pressure scaling
 ntf=1,            ! Complete force evaluation
 ntwx= 1000,       ! Write to trajectory file every ntwx steps
 ntpr= 1000,       ! Print to mdout every ntpr steps
 ntwr= 1000,       ! Write a restart file every ntwr steps
 cut=  8.0,        ! Nonbonded cutoff in Angstroms
 ntr=1,            ! Turn on restraints
 restraintmask=":1-350 & !@H=", ! atoms to be restrained
 restraint_wt=5.0, ! force constant for restraint
 ntxo=1,           ! Write coordinate file in ASCII format
 ioutfm=0,         ! Write trajectory file in ASCII format
 /

Create a file 02.equil.mdin and add the following lines:

 MD simualation
 &cntrl
 imin=0,           ! Perform MD
 nstlim=50000      ! Number of MD steps
 ntb=2,            ! Constant Pressure
 ntc=1,            ! No SHAKE on bonds between hydrogens
 dt=0.001,         ! Timestep (ps)
 ntp=1,            ! Isotropic pressure scaling
 barostat=1        ! Berendsen
 taup=0.5          ! Pressure relaxtion time (ps)
 ntf=1,            ! Complete force evaluation
 ntt=3,            ! Langevin thermostat
 gamma_ln=2.0      ! Collision Frequency for thermostat
 ig=-1,            ! Random seed for thermostat
 temp0=298.15      ! Simulation temperature (K)
 ntwx= 1000,       ! Write to trajectory file every ntwx steps
 ntpr= 1000,       ! Print to mdout every ntpr steps
 ntwr= 1000,       ! Write a restart file every ntwr steps
 cut=  8.0,        ! Nonbonded cutoff in Angstroms
 ntr=1,            ! Turn on restraints
 restraintmask=":1-235 & !@H=", ! atoms to be restrained
 restraint_wt=5.0, ! force constant for restraint
 ntxo=1,           ! Write coordinate file in ASCII format
 ioutfm=0,         ! Write trajectory file in ASCII format
 iwrap=1,          ! iwrap is turned on
 /

Create a file 03.min.mdin and add the following lines:

Minmize all the hydrogens
 &cntrl
 imin=1,           ! Minimize the initial structure
 maxcyc=1000,    ! Maximum number of cycles for minimization
 ntb=1,            ! Constant volume
 ntp=0,            ! No pressure scaling
 ntf=1,            ! Complete force evaluation
 ntwx= 1000,       ! Write to trajectory file every ntwx steps
 ntpr= 1000,       ! Print to mdout every ntpr steps
 ntwr= 1000,       ! Write a restart file every ntwr steps
 cut=  8.0,        ! Nonbonded cutoff in Angstroms
 ntr=1,            ! Turn on restraints
 restraintmask=":1-350 & !@H=", ! atoms to be restrained
 restraint_wt=2.0, ! force constant for restraint
 ntxo=1,           ! Write coordinate file in ASCII format
 ioutfm=0,         ! Write trajectory file in ASCII format
 /

Create a file 04.min.mdin and add the following lines:

Minmize all the hydrogens
&cntrl
imin=1,           ! Minimize the initial structure
maxcyc=1000,    ! Maximum number of cycles for minimization
ntb=1,            ! Constant volume
ntp=0,            ! No pressure scaling
ntf=1,            ! Complete force evaluation
ntwx= 1000,       ! Write to trajectory file every ntwx steps
ntpr= 1000,       ! Print to mdout every ntpr steps
ntwr= 1000,       ! Write a restart file every ntwr steps
cut=  8.0,        ! Nonbonded cutoff in Angstroms
ntr=1,            ! Turn on restraints
restraintmask=":1-350 & !@H=", ! atoms to be restrained
restraint_wt=0.1, ! force constant for restraint
ntxo=1,           ! Write coordinate file in ASCII format
ioutfm=0,         ! Write trajectory file in ASCII format
/

Create a file 05.min.mdin and add the following lines:

Minmize all the hydrogens
&cntrl
imin=1,           ! Minimize the initial structure
maxcyc=1000,    ! Maximum number of cycles for minimization
ntb=1,            ! Constant volume
ntp=0,            ! No pressure scaling
ntf=1,            ! Complete force evaluation
ntwx= 1000,       ! Write to trajectory file every ntwx steps
ntpr= 1000,       ! Print to mdout every ntpr steps
ntwr= 1000,       ! Write a restart file every ntwr steps
cut=  8.0,        ! Nonbonded cutoff in Angstroms
ntr=1,            ! Turn on restraints
restraintmask=":1-350 & !@H=", ! atoms to be restrained
restraint_wt=0.05, ! force constant for restraint
ntxo=1,           ! Write coordinate file in ASCII format
ioutfm=0,         ! Write trajectory file in ASCII format
/

Create a file 06.equil.mdin and add the following lines:

MD simualation
&cntrl
imin=0,           ! Perform MD
nstlim=50000      ! Number of MD steps
ntb=2,            ! Constant Pressure
ntc=1,            ! No SHAKE on bonds between hydrogens
dt=0.001,         ! Timestep (ps)
ntp=1,            ! Isotropic pressure scaling
barostat=1        ! Berendsen
taup=0.5          ! Pressure relaxtion time (ps)
ntf=1,            ! Complete force evaluation
ntt=3,            ! Langevin thermostat
gamma_ln=2.0      ! Collision Frequency for thermostat
ig=-1,            ! Random seed for thermostat
temp0=298.15      ! Simulation temperature (K)
ntwx= 1000,       ! Write to trajectory file every ntwx steps
ntpr= 1000,       ! Print to mdout every ntpr steps
ntwr= 1000,       ! Write a restart file every ntwr steps
cut=  8.0,        ! Nonbonded cutoff in Angstroms
ntr=1,            ! Turn on restraints
restraintmask=":1-350 & !@H=", ! atoms to be restrained
restraint_wt=1.0, ! force constant for restraint
ntxo=1,           ! Write coordinate file in ASCII format
ioutfm=0,         ! Write trajectory file in ASCII format
iwrap=1,          ! iwrap is turned on
/

Create a file 07.equil.mdin and add the following lines:

MD simulation
&cntrl
imin=0,           ! Perform MD
nstlim=50000      ! Number of MD steps
ntx=5,            ! Positions and velocities read formatted
irest=1,          ! Restart calculation
ntc=1,            ! No SHAKE on for bonds with hydrogen
dt=0.001,         ! Timestep (ps)
ntb=2,            ! Constant Pressure
ntp=1,            ! Isotropic pressure scaling
barostat=1        ! Berendsen
taup=0.5          ! Pressure relaxtion time (ps)
ntf=1,            ! Complete force evaluation
ntt=3,            ! Langevin thermostat
gamma_ln=2.0      ! Collision Frequency for thermostat
ig=-1,            ! Random seed for thermostat
temp0=298.15      ! Simulation temperature (K)
ntwx= 1000,       ! Write to trajectory file every ntwx steps
ntpr= 1000,       ! Print to mdout every ntpr steps
ntwr= 1000,       ! Write a restart file every ntwr steps
cut=  8.0,        ! Nonbonded cutoff in Angstroms
ntr=1,            ! Turn on restraints
restraintmask=":1-350 & !@H=", ! atoms to be restrained
restraint_wt=0.5, ! force constant for restraint
ntxo=1,           ! Write coordinate file in ASCII format
ioutfm=0,         ! Write trajectory file in ASCII format
iwrap=1,          ! iwrap is turned on
/

Create a file 08.equil.mdin and add the following lines:

MD simulations
&cntrl
imin=0,           ! Perform MD
nstlim=50000      ! Number of MD steps
ntx=5,            ! Positions and velocities read formatted
irest=1,          ! Restart calculation
ntc=1,            ! No SHAKE on for bonds with hydrogen
dt=0.001,         ! Timestep (ps)
ntb=2,            ! Constant Pressure
ntp=1,            ! Isotropic pressure scaling
barostat=1        ! Berendsen
taup=0.5          ! Pressure relaxtion time (ps)
ntf=1,            ! Complete force evaluation
ntt=3,            ! Langevin thermostat
gamma_ln=2.0      ! Collision Frequency for thermostat
ig=-1,            ! Random seed for thermostat
temp0=298.15      ! Simulation temperature (K)
ntwx= 1000,       ! Write to trajectory file every ntwx steps
ntpr= 1000,       ! Print to mdout every ntpr steps
ntwr= 1000,       ! Write a restart file every ntwr steps
cut=  8.0,        ! Nonbonded cutoff in Angstroms
ntr=1,            ! Turn on restraints
restraintmask=":1-349@CA.C.N", ! atoms to be restrained, only the backbone
restraint_wt=0.1, ! force constant for restraint
ntxo=1,           ! Write coordinate file in ASCII format
ioutfm=0,         ! Write trajectory file in ASCII format
iwrap=1,          ! iwrap is turned on
/

Create a file 09.equil.mdin and add the following lines:

MD simulations
&cntrl
imin=0,           ! Perform MD
nstlim=50000      ! Number of MD steps
ntx=5,            ! Positions and velocities read formatted
irest=1,          ! Restart calculation
ntc=1,            ! No SHAKE on for bonds with hydrogen
dt=0.001,         ! Timestep (ps)
ntb=2,            ! Constant Pressure
ntp=1,            ! Isotropic pressure scaling
barostat=1        ! Berendsen
taup=0.5          ! Pressure relaxtion time (ps)
ntf=1,            ! Complete force evaluation
ntt=3,            ! Langevin thermostat
gamma_ln=2.0      ! Collision Frequency for thermostat
ig=-1,            ! Random seed for thermostat
temp0=298.15      ! Simulation temperature (K)
ntwx= 1000,       ! Write to trajectory file every ntwx steps
ntpr= 1000,       ! Print to mdout every ntpr steps
ntwr= 1000,       ! Write a restart file every ntwr steps
cut=  8.0,        ! Nonbonded cutoff in Angstroms
ntr=1,            ! Turn on restraints
restraintmask=":1-349@CA.C.N", ! atoms to be restrained
restraint_wt=0.1, ! force constant for restraint
ntxo=1,           ! Write coordinate file in ASCII format
ioutfm=0,         ! Write trajectory file in ASCII format
iwrap=1,          ! iwrap is turned on
/

This should not be run on the head node, instead create a submission script called equil.sh and add the following lines:

#! /bin/sh
#SBATCH --job-name=sys_equilibration
#SBATCH --ntasks-per-node=40
#SBATCH --nodes=2
#SBATCH --time=8:00:00
#SBATCH -p long-40core
cd $SLURM_SUBMIT_DIR
echo "Started Equilibration on `date` "
do_parallel="mpirun pmemd.MPI"
prmtop="../001.tleap_build/2nnq.wet.complex.prmtop"
coords="../001.tleap_build/2nnq.wet.complex"
MDINPUTS=(01.min 02.equil 03.min 04.min 05.min 06.equil 07.equil 08.equil 09.equil)
for input in ${MDINPUTS[@]}; do
$do_parallel -O -i ${input}.mdin -o ${input}.mdout -p $prmtop -c ${coords}.rst7 -ref ${coords}.rst7 -x ${input}.trj -inf ${input}.info -r 
${input}.rst7
coords=$input
done
echo "Finished Equilibration on `date` "

Finally run equilibrations using:

sbatch equil.sh

This simulations will take some time. Once all steps have completed, be sure to check the output.txt files and ensure they are reasonable.

Production

Now the system is ready for production. Production runs can be run with restrains on the protein, or without restraints on the protein. In this tutorial we will keep the restrains on the protein but this can be changed by removing the "restraintmask" and "restraint_wt" variables. Now navigate to the 003.production directory.

Create a file 10.prod.mdin and add the following lines:

MD simulations
 &cntrl
  imin=0,           ! Perform MD
  nstlim=5000000,   ! Number of MD steps
  ntx=5,            ! Positions and velocities read formatted
  irest=1,          ! Restart calculation
  ntc=2,            ! SHAKE on for bonds with hydrogen
  dt=0.002,         ! Timestep (ps)
  ntb=2,            ! Constant Pressure
  ntp=1,            ! Isotropic pressure scaling
  barostat=1        ! Berendsen
  taup=0.5          ! Pressure relaxtion time (ps)
  ntf=2,            ! No force evaluation for bonds with hydrogen
  ntt=3,            ! Langevin thermostat
  gamma_ln=2.0      ! Collision Frequency for thermostat
  ig=-1,            ! Random seed for thermostat
  temp0=298.15      ! Simulation temperature (K)
  ntwx= 2500,       ! Write to trajectory file every ntwx steps
  ntpr= 2500,       ! Print to mdout every ntpr steps
  ntwr= 5000000,    ! Write a restart file every ntwr steps
  cut=8.0,          ! Nonbonded cutoff in Angstroms
  ntr=1,            ! Turn on restraints
  restraintmask=":1-349@CA,C,N", ! atoms to be restrained
  restraint_wt=0.1, ! force constant for restraint
  ntxo=1,           ! Write coordinate file in ASCII format
  ioutfm=0,         ! Write trajectory file in ASCII format
  iwrap=1,          ! iwrap is turned on
 $end

This simulation will also require a slurm script. This will take a very long time so make sure you select a queue that will allow it to finish.

Create a file prod.sh and add the following lines:

#! /bin/sh
#SBATCH --job-name=production_unrest
#SBATCH --ntasks-per-node=28
#SBATCH --nodes=1
#SBATCH --time=7-00:00:0
#SBATCH -p extended-28core
cd $SLURM_SUBMIT_DIR
echo "Started Production on `date` "
#do_parallel="mpirun pmemd.MPI"
do_parallel="/gpfs/software/intel/parallel-studio-xe/2018_3/compilers_and_libraries/linux/mpi/bin64/mpirun -np 80 /gpfs/software/amber/16/intel/cpu/bin/pmemd.MPI"
#do_cuda="pmemd.cuda"
prmtop="../../001.tleap_build/1ndv.wet.complex.prmtop"
coords="../../002.equilbration/09.equil"
MDINPUTS=(10.prod)
for input in ${MDINPUTS[@]}; do
$do_parallel -O -i ${input}.mdin -o ${input}.mdout -p $prmtop -c ${coords}.rst7 -ref ${coords}.rst7 -x ${input}.trj -inf ${input}.info -r 
${input}.rst7
 coords=$input
done
echo "Finished Production on `date` "

Now submit your job using:

sbatch prod.sh

Once production is finished you should see a "10.prod.mdout" and a "10.prod.trj". View the "10.prod.mdout" file to ensure that the simulation ran to completion.

Analysis

RMSD

The first analysis we will be performing is an RMSD analysis. This will tell us how much the ligand, protein, or complex moved from its initial position during simulation. We will also be creating a trajectory file with only the protein and ligand atoms. This will be useful if you wanted to view the trajectory in Chimera or another software.

To make the stripped trajectory and calculate RMSD we will be using a tool called CPPTRAJ. Create a file called cpptraj_strip_wat.in and add the following lines:

#!/usr/bin/sh
parm ../001.tleap_build/1ndv.wet.complex.parm7
#read in trajectory
trajin ../003.production/10.prod.trj
#strip solvent
strip :WAT:Na+:Cl-
#create gas-phase trajectory
trajout 1ndv_gas.trj nobox

Run this file using the following command:

cpptraj -i cpptraj_strip_wat.in

The .trj generating will be smaller and easier to load moving forward. Now lets create a new input file to calculate the RMSD. Create a file called cpptraj_rmsd.in and add the following lines:

#!/usr/bin/sh
parm ../001.leap_build/1ndv.gas.complex.parm7
#trajin the trajectory
trajin 1ndv_gas.trj
#read in reference
reference ../001.tleap_build/3wze.gas.complex.rst7
#compute RMSD (do not fit internal geometris first, included rigid body motions, convert frames to ns (framenum*.005)
rmsd rms1 ":350&!(@H=)" nofit mass out 1ndv_lig_restrained_rmsd_nofit.dat time .005
#histogram the nofit rmsd
histogram rms1,*,*,.1,* norm out 1ndv_lig_restrained_rmsd_nofit_histogram.dat
rmsd rms2 ":1-349&!(@H=)" nofit mass out 1ndv_protein_restrained_rmsd_nofit.dat time .005
#histogram the nofit rmsd
histogram rms1,*,*,.1,* norm out 1ndv_protein_restrained_rmsd_nofit_histogram.dat

Run the file using the command:

cpptraj -i cpptraj_rmsd.in

This file loads in our paramter file for the gas complex, followed by our trajectory and our reference of the first frame. It then computes the RMSD of just the ligand, and then just the protein as well as histogram data for them. These files can then be graphed.

Hydrogen Bonding

We will now analyze the hydrogen bond formed between the ligand and the protein. We will gain be using CPPTRAJ to do this. Note that this script will tell us all the hydrogen bonds that are formed and how long they persist in simulation, not just the ones between the ligand and the protein. Create a file called cpptraj_hbond.in and add the following lines:

#!/usr/bin/sh
parm ../001.leap_build/1ndv.gas.complex.parm7
#trajin the trajectory
trajin 1ndv_gas.trj
#read in reference
#wrap everything in one periodic cell - could cause problems, may comment out #autoimage if problems later
autoimage
#compute intra + water-mediated H-bonds
hbond hb1 :1-350 out 1ndv_hbond.out avgout 1ndv_hbond_avg.dat solventdonor :WAT solventacceptor :WAT@-O nointramol bridgeout 1ndv_bridge-water.dat dist 3.0 angle 140
hbond hb2 :1-350 out 1ndv_sunitinib.hbond.out avgout 1ndv_sunitinib.hbond.avg.dat solventdonor :WAT solventacceptor :WAT nointramol bridgeout 1ndv_sunitinib.bridge-water.dat dist 3.0 angle 140

Run the script using the following command:

cpptraj -i cpptraj_hbond.in

MM-GBSA

The last analysis we will perform is an MM-GBSA calculation to estimate the free energy of binding. Create the file mmgbsa.in and add the following lines:

mmgbsa 1ndv analysis
&general
 interval=1, netcdf=1,
 keep_files=0,
/
&gb
 igb=8
 saltcon=0.0, surften=0.0072,
 surfoff=0.0, molsurf=0,
/
&nmode
 drms=0.001, maxcyc=10000,
 nminterval=250, nmendframe=2000,
 nmode_igb=1,
/

This is a complex calcualtion and should not be run on the head node. Create a submissions script called mmgbsa.sh and add the following lines:

#! /bin/sh
#SBATCH --job-name=production_unrest
#SBATCH --ntasks-per-node=28
#SBATCH --nodes=1
#SBATCH --time=7-00:00:0
#SBATCH -p extended-28core
cd $SLURM_SUBMIT_DIR

solv_parm="../001.tleap_build/1ndv.wet.complex.prmtop" complex_parm="../001.tleap_build/1ndv.gas.complex.parm7" receptor_parm="../001.tleap_build/1ndv.gas.receptor.parm7" lig_parm="../001.tleap_build/1ndv.gas.ligand.parm7" trajectory="../003.production/10_prod.trj" MMPBSA.py -O -i mmgbsa.in \

    -o  1ndv_mmgbsa_results.dat \ 
    -eo 1ndv_mmgbsa_perframe.dat \
    -sp ${solv_parm} \
    -cp ${complex_parm} \
    -rp ${receptor_parm} \
    -lp ${lig_parm} \
     -y ${trajectory}

Difference between revisions of "2024 AMBER tutorial 2 with PDBID 1NDV"

Latest revision as of 22:22, 5 May 2024

Contents

Introduction

Getting Started

Before Simulation

Structure

Force Field Parameters

TLEap

Minimization and Equilibration

Production

Analysis

RMSD

Hydrogen Bonding

MM-GBSA

Navigation menu

Personal tools

Namespaces

Variants

Views

More

Search

Navigation

Rizzo Lab

Courses

Toolbox