Here we give a brief overview of the necessary steps to run the program but we strongly encourage to follow the tutorials. The user guide is partitioned in three levels of usage:
For flexible fitting a single atomic structure into a EM density map enter the following command at the prompt.
imodfit <pdb> <map> <resolution> <cutoff>
where:
pdb | PDB initial model (required). |
map | Target EM map file (required). Situs and CCP4 formats are accepted; but extensions ".sit" and ".cpp4" are mandatory. |
resolution | Resolution (required). Our resolution criterion follows EMAN package procedures (e.g. PDB2MRC tool) |
Cutoff | EM map threshold (required). All density levels below this value will not be considered. |
iMODFIT outputs the following files:
- imodfit_fitted.pdb --> fitted atomic structure
- imodfit_score.txt --> score file to check for convergence
- imodfit.log--> used command log
The more common options to customize your fitting are:
−h | Displays usage information and exits |
−i <int> | Maximum number of iterations (default=10000). |
−m | Coarse-Grained model: 0=CA, 1=3BB2R, 2=Full-Atom, 3=NCAC(experimental) (default=2). |
−x | Considers first CHI dihedral angle (default=disabled). |
−t | Outputs a Multi-PDB trajectory movie (<basename_movie>.pdb) |
−n <int/float> | Used modes range, either number [1,N] <integer>, or ratio [0,1) <float> (default=0.2). |
−e <int/float> | Excited modes range, either number [1,nevs] <integer>, or ratio [0,1) <float> (default=0.02) |
−F | Enables full-atom output models. |
−o <string> | Output files basename (default=imodfit). |
−P <int> | Pairwise interaction potential: (default=0) 0=inverse exponential (= k/(1+(x/x0)^p), if x < c, else k=0) 1=cutoff (= k, if x < c, else k=0) 2=Topology & Secondary Structure (--func is mandatory) By default an extra torsional potential will be added. |
−f <string> | ASCII file defining the ICs to be fixed with the format: Protein: "index phi chi psi" NAcid: "index alpha beta gamma chi epsilon zeta" Inter-chain: "index IC" Where IC-name= 0(fixed) or 1(mobile) A demo file can be generated using iMODE with the --save_fixfile option. |
−r <float> | Randomly fixed ratio of Dihedral Coords. Example: 0.7 = 70% of dihedrals will be randomly fixed. (Rotational/Translational coords. always mobile) |
−S | All dihedral coordinates belonging to residues matching the indicated SS identifieres will be fixed. Ex: "HE" will fix the dihedrals corresponding to alpha-helices and beta-sheets. |
Only for real expert users!
--ss <string> Secondary Structure ASCII file with 2 cols.: <index> <char> Where <index> is the corresponding residue index (1,2,...), and <char> is the single character SS identifier. By default SS will be computed internally (H=helix, E=strand, C=coil). --func <string> ASCII file defining the force constant functions to be applied according to Topology and/or Secondary Structure. The 5 cols. format is: <SS> <n> <k> <x0> <pow> Where <SS> is the two character pairwise interaction identifier, <n> is the topology, and <k>,<x0>,<pow> are the corresponding inverse exponential function parameters. If --ss is not specified, only the XX pairwise interaction identifier will be considered. If <n> is 0, any previously not-matched topology will be matched. --k0_c <float> Inverse Exponential's distance cutoff (default=10A) --k0_k <float> Inverse Exponential's stiffness constant (default=1.0) --k0_x0 <float> Inverse Exponential's inflexion point (default=3.8A) --k0_p <float> Inverse Exponential's power term (default=6) --k1_c <float> Distance cutoff method distance cutoff (default=10A) --k1_k <float> Distance cutoff method stiffness constant (default=1.0) --k2_c <float> Non-bonding distance cutoff applied to --ss_func (default=10A) --pdb_ref <string> Reference PDB file (test only) --morepdbs Enable initial (basename_model.pdb) pdb model output (default=disabled) --nomass Disables mass weighting (default=disabled) --notors Disables extra torsional potential (default=disabled) --nowrmsd Disables Gaussian Weighted RMSD (default=disabled) --cutoff2 <float> Density cutoff of simulated map (default = 0.001) --filter <int> Select filtration method: 1-Fourier, 2-Kernel (By default the fastest will be selected) --6Dref <int> Number of iterations between local 6D pose refinement (default=200) --6Dref_delay <int> Number of iterations before first local 6D pose refinement (default=disabled) --6Dref_chain Independent local 6D pose refinement for each single chain. --6Dref_trans <float> Maximum translational increment (Angstroms) for pose refinement. (default=2) --6Dref_rot <float> Maximum rotational increment (degrees) for pose refinement (default=1) --prob <string> Normal Mode Selection Probabitity. --nex modes will be selected and merged from --nevs subset according to the following probabilities (p): (default=var) plain: equiprobability (--min_prob will be ignored) var: proportional to the mode variance, p(i)= 1/eigenvalue(i) line: lineally decreasing probability, p(i)= 1-i/nevs --rediag <double> RMSD ratio to trigger diagonalization. (last_diag_RMSD-current_RMSD)/first_RMSD > rediag (default=0.05) --delta_save <float> RMSD increment from previous saved frame to trigger frame saving. (default=0.5) --wrmsd <float> Sets the RMSD weighting [0:1] during model alingment (wrmsd=0, means maximum weighting) (default=0) --conv <double> Convergence threshold (relative to first score) (default=1e-8) --conv_win <int> Window length (number of iterations) to estimate convergence. (default=5000) --seed <unsigned int> Pre-define the random number generator SEED (Mersenne Twister) (default=random-seed from /dev/urandom) --verb <int> Verbosity display level (0=low, 1=medium, 2=high) (default=0) --step <double> Initial amplitude applied to excited modes (default=10) --step2 <double> Final amplitude applied to excited modes (amplitude will decrease lineally from "step" to "step2") (default=step/10) --rand_step Random amplitude selection between step and step2 (default=disabled) --nex2 <int> Final number of excited eigenvectors, either number [1,nevs] <integer>, or ratio [0,1) <float>. Number of excited modes will change lineally from "nevs" to "nevs2" (default=disabled) --rand_excited Random number of excited modes between nex and nex2 (default=disabled) --norand_weight Disables random excited modes weighting (default=disabled) --, --ignore_rest Ignores the rest of the labeled arguments following this flag. -v, --version Displays version information and exits.