Structure calculation using manually assigned NOESY peak lists

This calculation shows how to calculate a structure from previously assigned NOESY peak lists. The input data are in the subdirectory ‘manual’ of the demo data zip archive:

demo.seq

amino acid sequence

c13.peaks

peak list from 3D ¹³C-resolved NOESY spectrum

n15.peaks

peak list from 3D ¹⁵N-resolved NOESY spectrum

aro.peaks

peak list from 3D aromatic ¹³N-resolved NOESY spectrum

demo.prot

¹H, ¹³C, and ¹⁵N chemical shift list

demo.aco

dihedral angle restraints

init.cya

initialization macro

CALC.cya

macro for the structure calculation

The structure calculation is performed by the commands in the macro file CALC.cya:

name       := demo
peaks      := c13,n15,aro                  # names of peak list(s) 
prot       := $name                        # names of proton list(s)
tolerance  := 0.040,0.030,0.45             # shift tolerances: H, H', C/N', C/N
calibration:= 6.7E5,8.2E5,8.0E4            # calibration constants, automatic if empty
dref       := 4.2                          # average distance limit for automatic calibration

if (master) then                           # execute the following commands only on master node

  # ---- check consistency of peak and chemical shift lists----

  peakcheck peaks=$peaks prot=$prot        # check consistency of peak and shift assignments

  # ---- calibration ----

  calibration prot=$prot peaks=$peaks constant=$val('calibration') dref=$dref  # calibrate NOEs
  peaks calibrate "**" simple              # convert calibrated NOEs into distance restraints
  write upl $name-in.upl                   # write initial upper distance limit file
  distance modify                          # remove irrelevant restraints, apply pseudoatom corrections
  write upl $name.upl                      # write upper distance limit file

end if
synchronize                                # wait until all parallel processes have arrived here

# ---- structure calculation ----

read seq $name.seq                         # re-read sequence to initialize
read upl $name.upl                         # read upper distance limits
read aco $name.aco                         # read angle restraints
seed=5671                                  # set random number generator seed
calc_all structures=100 command=anneal steps=10000   # calculate conformers
overview $name.ovw structures=20 pdb       # write overview file and coordinates

First several variables are set: The variable name is used to define the name of various input and output files. The variable peaks gives the names of the input peak lists, separated by commas without intervening blanks. The variable prot gives the name(s) of the input chemical shift list(s). If a single name is given as in the example, it specifies that a single chemical shift list file with this name will be used for all peak lists. Alternatively, it is possible to specify a separate chemical shift list for each peak list as a comma-separated list of file names. The variable tolerance specifies The first two commands read the input files with upper distance bounds and dihedral angle restraints, respectively. Then, 50 conformers are calculated using the standard simulated annealing schedule with 4000 torsion angle dynamics steps per conformer. Finally, the 10 conformers with the lowest final target function values are analyzed. An overview table of these 10 best conformers is saved in the file demo.ovw, and their coordinates are written to the PDB file demo.pdb.