[gmx-users] fep protocol for gromacs 2016

Yoav Atsmon-Raz yoav.atsmonraz at ucalgary.ca
Tue Jan 9 06:19:40 CET 2018


Hi everyone,

I’ve been trying to get a the free energy difference between a leucine residue vs an alanine residue by doing a set of FEP simulations in which I decouple the VDW and electrostatic interactions in the martini CG force field. In the particular example I’m using here, I’m aiming to decouple the Leucine sidechain and the topology I’m using is as follows:
--------------------------------------
[ moleculetype ]
; Name         Exclusions
Protein_A         1

[ atoms ]
    1    P5     1   TRP    BB     1  0.0000 P5   0  72
    2   SC4     1   TRP   SC1     2  0.0000 SC4  0  72
    3   SNd     1   TRP   SC2     3  0.0000 SNd  0  72
    4   SC5     1   TRP   SC3     4  0.0000 SC5  0  72
    5   SC5     1   TRP   SC4     5  0.0000 SC5  0  72
    6   Nda     2   LEU    BB     6  0.0000 Nda  0  72
    7   AC1     2   LEU   SC1     7  0.0000 AC1  0  72
    8   Nda     3   LEU    BB     8  0.0000 Nda  0  72
    9   AC1     3   LEU   SC1     9  0.0000 DUM  0  72
   10   Nda     4   LEU    BB    10  0.0000 Nda  0  72
   11   AC1     4   LEU   SC1    11  0.0000 AC1  0  72
   12    Qa     5   LEU    BB    12 -1.0000 Qa  -1  72
   13   AC1     5   LEU   SC1    13  0.0000 AC1  0  72

[ bonds ]
; Backbone bonds
    1     6      1   0.35000  1250 ; TRP(E)-LEU(E)
    6     8      1   0.35000  1250 ; LEU(E)-LEU(E)
    8    10      1   0.35000  1250 ; LEU(E)-LEU(E)
   10    12      1   0.35000  1250 ; LEU(E)-LEU(E)
; Sidechain bonds
    1     2      1   0.30000  5000 ; TRP
    6     7      1   0.33000  7500 ; LEU
    8     9      1   0.33000  7500 ; LEU
   10    11      1   0.33000  7500 ; LEU
   12    13      1   0.33000  7500 ; LEU
; Short elastic bonds for extended regions
    1     8      1   0.64000  2500 ; TRP(1)-LEU(8) 1-3
    6    10      1   0.64000  2500 ; LEU(6)-LEU(10) 2-4
    8    12      1   0.64000  2500 ; LEU(8)-LEU(12) 2-4
; Long elastic bonds for extended regions
    1    10      1   0.97000  2500 ; TRP(1)-LEU(10) 1-4
    6    12      1   0.97000  2500 ; LEU(6)-LEU(12) 1-4

[ constraints ]
    2     3      1   0.27000 ; TRP
    2     4      1   0.27000 ; TRP
    3     4      1   0.27000 ; TRP
    3     5      1   0.27000 ; TRP
    4     5      1   0.27000 ; TRP
[ angles ]
; Backbone angles
    1     6     8      2    134    25 ; TRP(E)-LEU(E)-LEU(E)
    6     8    10      2    134    25 ; LEU(E)-LEU(E)-LEU(E)
    8    10    12      2    134    25 ; LEU(E)-LEU(E)-LEU(E)
; Backbone-sidechain angles
    2     1     6      2    100    25 ; TRP(E)-LEU(E) SBB
    1     6     7      2    100    25 ; TRP(E)-LEU(E) SBB
    6     8     9      2    100    25 ; LEU(E)-LEU(E) SBB
    8    10    11      2    100    25 ; LEU(E)-LEU(E) SBB
   10    12    13      2    100    25 ; LEU(E)-LEU(E) SBB
; Sidechain angles
    1     2     3      2    210    50 ; TRP
    1     2     4      2     90    50 ; TRP

[ dihedrals ]
; Backbone dihedrals
; Sidechain improper dihedrals
    1     3     4     2      2      0    50 ; TRP
    2     3     5     4      2      0   200 ; TRP
-------------------------------------------


The mdp I’m using goes like this:

-----------------------------------------
; Run control
integrator               = sd       ; Langevin dynamics
tinit                    = 0
dt                       = 0.02
nsteps                   = 50000000   ; 1 us
nstcomm                  = 100
comm-grps                = Protein_A Water
; Output control
nstxout                  = 0
nstvout                  = 0
nstfout                  = 0
nstlog                   = 1000
nstenergy                = 100
nstxout-compressed       = 1000
compressed-x-precision   = 10
compressed-x-grps        = Protein_A Water
energygrps               = Protein_A Water
; Neighborsearching and short-range nonbonded interactions
cutoff-scheme            = verlet
verlet-buffer-tolerance  = 0.005
nstlist                  = 20
ns_type                  = grid
pbc                      = xyz
; Electrostatics
coulombtype              = reaction-field
rcoulomb                 = 1.1
epsilon_r                = 15
epsilon_rf               = 0
; van der Waals
vdwtype                  = cutoff
vdw-modifier             = Potential-shift-verlet
rvdw                     = 1.1
; Temperature coupling
; tcoupl is implicitly handled by the sd integrator
tc_grps                  = Protein_A Water
tau_t                    = 1.0 1.0
ref_t                    = 300 300
; Pressure coupling is on for NPT
Pcoupl                   = Berendsen
Pcoupltype               = isotropic
tau_p                    =  5.0
compressibility          = 3e-04 3e-04
ref_p                    = 1.0 1.0
; Free energy control stuff
free_energy              = yes
init_lambda_state        = 0
delta_lambda             = 0
calc_lambda_neighbors    = -1        ; only immediate neighboring windows
; Vectors of lambda specified here
; Each combination is an index that is retrieved from init_lambda_state for each simulation
; init_lambda_state        0    1    2    3    4    5    6    7    8    9    10   11   12   13   14   15   16   17   18   19   20   21   22   23   24   25
vdw_lambdas              = 0.00 0.00 0.00 0.00 0.00 0.00 0.05 0.10 0.15 0.20 0.25 0.30 0.35 0.40 0.45 0.50 0.55 0.60 0.65 0.70 0.75 0.80 0.85 0.90 0.95 1.00
coul_lambdas             = 0.00 0.20 0.40 0.60 0.80 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00
; We are not transforming any bonded or restrained interactions
bonded_lambdas           = 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00
restraint_lambdas        = 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00
; Masses are not changing (particle identities are the same at lambda = 0 and lambda = 1)
mass_lambdas             = 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00
; Not doing simulated temperting here
temperature_lambdas      = 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00
; Options for the decoupling
sc-alpha                 = 0.5
sc-coul                  = no       ; linear interpolation of Coulomb (none in this case)
sc-power                 = 1
sc-sigma                 = 0.3
couple-moltype           = Protein_A  ; name of moleculetype to decouple
couple-lambda0           = vdw-q      ; van der Waals interactions and coul
couple-lambda1           = none ; turn off everything, in this case only vdW
couple-intramol          = yes
nstdhdl                  = 100
; Do not generate velocities
gen_vel                  = no
gen_temp                 = 300
gen_seed                 = 473529
; options for bonds
constraints              = none  ; we only have C-H bonds here
; Type of constraint algorithm
constraint-algorithm     = lincs
; Constrain the starting configuration
; since we are continuing from NPT
continuation             = yes
; Highest order in the expansion of the constraint coupling matrix
lincs-order              = 12


Can someone just tell me if I’m doing this correctly? The values I’m getting via MBAR in alchemical analysis seem to high for me (~170 kj/mol) and I want to be sure that its currently indeed addressing the switch from topology A to the mutant topology B. Also I’m using GMX 2016.3

Thanks in advance for everyone, Yoav



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