Resonance Assignment/Abacus/Structure calculation setup and analysis: Difference between revisions
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=== Step 1. Load Data === | === Step 1. Load Data === | ||
<div> </div> | <div> </div> | ||
**open new project PRJ3 { | **open new project PRJ3 { [[FMCGUI_commands#Project.3ENew|Project>new]] } | ||
**load 15N NOESY peak list { [[ | **load 15N NOESY peak list { [[FMCGUI commands#Data.3EN15_NOESY.3E|DATA>N15 NOESY>load]] } | ||
**load 13C_aliphatic NOESY peak list { [[ | **load 13C_aliphatic NOESY peak list { [[FMCGUI commands#Data.3EC13_NOESY.3E|DATA>C13NOESY H2O>load]] } | ||
**load 13C_aromatic NOESY peak list { [[ | **load 13C_aromatic NOESY peak list { [[FMCGUI commands#Data.3EArom_NOESY.3E|DATA>AromNOESY>load]] } | ||
**set tolerances { [[ | **set tolerances { [[FMCGUI commands#Data.3ETolerances|Data>Tolerances]] } <span /> | ||
**<span>load ''<u>assigned </u>''chemical shifts { [[FMCGUI commands#Fragment.3ELoad.3Eassigned|Fragment>Load>assigned]] } | |||
**<span>load ''<u>assigned </u>''chemical shifts [Fragment>Load>assigned] </span> | </span> | ||
<div><span> The file with assigned chemical shifts could be either in “standard” format (assigned AA-fragments) or in cyana | <div><span> The file with assigned chemical shifts could be either in “standard” format ([[Spin systems format|assigned AA-fragments]]) or in [[Spin systems format|cyana format]] (prot-file).</span></div><div> </div><div><span> </span></div> | ||
=== Step 2. Set up constraints === | === Step 2. Set up constraints === | ||
<div> </div><div>''' '''The structure calculation requires dihedral angle constraints in the cyana format (aco-file). These constraints are usually prepared using the results of dihedral angle prediction by TALOS. H-bond constraints are optional.</div><div> </div> | <div> </div><div>''' '''The structure calculation requires dihedral angle constraints in the cyana format (aco-file). These constraints are usually prepared using the results of dihedral angle prediction by TALOS. H-bond constraints are optional.</div><div> </div> | ||
*<span><span> </span></span>calculate dihedral angle constraints [ | *<span><span> </span></span>calculate dihedral angle constraints { [[FMCGUI_commands#Structure.3EConstraints.3ETalos.3Ecalculate|Structure>Constraints>Talos>Calculate]] } | ||
*<span><span> </span></span>set up H-bond constraints [ | *<span><span> </span></span>set up H-bond constraints { [[FMCGUI_commands#Structure.3EConstraints.3EH-bonds.3ESpecify|Structure>Constraints>H-bonds>Specify]] } | ||
<div><span> </span></div><div>In the cased dihedral angle or H-bond constraints in cyana format (aco-file or upl-file, respectively) already are prepared, then the constraints can be loaded from the corresponding files <span> [Structure->Constraints->Talos>Load] or [Structure>Constraints>H-bonds>Load] </span></div><div> </div><div> </div> | <div><span> </span></div><div>In the cased dihedral angle or H-bond constraints in cyana format (aco-file or upl-file, respectively) already are prepared, then the constraints can be loaded from the corresponding files <span> { [[FMCGUI_commands#Structure.3EConstraints.3ETalos.3Eload|Structure->Constraints->Talos>Load]] } or { [[FMCGUI_commands#Structure.3EConstraints.3EH-bonds.3ESpecify|Structure>Constraints>H-bonds>Load]] } </span></div><div> </div><div> </div> | ||
=== Step 3. Specify ligands coordinating ZN ions (if there are any) === | |||
<div> </div><div>If there are ZN ions as a part of a protein structure the file “zn_ligands” should be present inside FMCGUI project directory. This file can be created by the following command</div><div>''' '''</div> | === Step 3. Specify ligands coordinating ZN ions (if there are any) === | ||
*<span> specify residues that coordinate ZN ion(s) [Structure>Add ZN ion] </span> | <div> </div><div>If there are ZN ions as a part of a protein structure the file “zn_ligands” should be present inside FMCGUI project directory. This file can be created by the following command</div><div>''' '''</div> | ||
*<span> specify residues that coordinate ZN ion(s) { [[FMCGUI commands#Structure.3EAdd_ZN_ligands|Structure>Add ZN ion]] } | |||
</span> | |||
<div>''' '''</div><div> </div> | <div>''' '''</div><div> </div> | ||
=== '''Step 4. Set up CYANA calculations''' === | |||
<div>''' '''</div> | === '''Step 4. Set up CYANA calculations''' === | ||
*<span> setting up structure calculations with CYANA [ | <div>''' '''</div> | ||
*<span> setting up structure calculations with CYANA { [[FMCGUI commands#Structure.3ECalculate.3ECyana|Structure>Calcuate>cyana]] } | |||
</span> | |||
<div>'''<span> </span>'''All files that are necessary for CYANA run are prepared and saved in the user specified directory, crun#, which is located inside the project directory. These files include chemical shifts (belok.prot file), sequence file, peak lists, dihedral angles constraints (file belok.aco), H-bond constraints, if available, (file hbond.upl), and constraints for ZN ions, if present, (files zn.upl, zn.lol).</div><div> </div> | <div>'''<span> </span>'''All files that are necessary for CYANA run are prepared and saved in the user specified directory, crun#, which is located inside the project directory. These files include chemical shifts (belok.prot file), sequence file, peak lists, dihedral angles constraints (file belok.aco), H-bond constraints, if available, (file hbond.upl), and constraints for ZN ions, if present, (files zn.upl, zn.lol).</div><div> </div> | ||
=== Step 5. Structure evaluation and peak lists refinement === | |||
<div> </div><div>RPF analysis and DP score allow one to estimate goodness-of-fit of a structural ensemble to NOESY peak lists. The results of RPF analysis can serve both for structure validation and peak lists refinement. </div><div> </div> | === Step 5. Structure evaluation and peak lists refinement === | ||
*<span><span> </span></span>run RPF analysis [ | <div> </div><div>RPF analysis and DP score allow one to estimate goodness-of-fit of a structural ensemble to NOESY peak lists. The results of RPF analysis can serve both for structure validation and peak lists refinement. </div><div> </div> | ||
*<span><span> </span></span>set up DP-score calculations with AutoStructure [ | *<span><span> </span></span>run RPF analysis { [[FMCGUI commands#Structure.3ERPF.3ERP|Structure>RPF>RP]] }. The results of the RPF analysis include peak lists in the SPARKY format of both false negative and false positive peaks for C13-aliphatic NOESY, C13-Aromatic NOESY, and N15-NOESY spectra in separate files. | ||
*<span><span> </span></span>set up DP-score calculations with AutoStructure { [[FMCGUI commands#Structure.3ERPF.3EDP|Structure>RPF>DP]] }. | |||
Latest revision as of 23:49, 5 January 2010
Step 1. Load Data
- open new project PRJ3 { Project>new }
- load 15N NOESY peak list { DATA>N15 NOESY>load }
- load 13C_aliphatic NOESY peak list { DATA>C13NOESY H2O>load }
- load 13C_aromatic NOESY peak list { DATA>AromNOESY>load }
- set tolerances { Data>Tolerances }
- load assigned chemical shifts { Fragment>Load>assigned }
The file with assigned chemical shifts could be either in “standard” format (assigned AA-fragments) or in cyana format (prot-file).
Step 2. Set up constraints
The structure calculation requires dihedral angle constraints in the cyana format (aco-file). These constraints are usually prepared using the results of dihedral angle prediction by TALOS. H-bond constraints are optional.
- calculate dihedral angle constraints { Structure>Constraints>Talos>Calculate }
- set up H-bond constraints { Structure>Constraints>H-bonds>Specify }
In the cased dihedral angle or H-bond constraints in cyana format (aco-file or upl-file, respectively) already are prepared, then the constraints can be loaded from the corresponding files { Structure->Constraints->Talos>Load } or { Structure>Constraints>H-bonds>Load }
Step 3. Specify ligands coordinating ZN ions (if there are any)
If there are ZN ions as a part of a protein structure the file “zn_ligands” should be present inside FMCGUI project directory. This file can be created by the following command
- specify residues that coordinate ZN ion(s) { Structure>Add ZN ion }
Step 4. Set up CYANA calculations
- setting up structure calculations with CYANA { Structure>Calcuate>cyana }
All files that are necessary for CYANA run are prepared and saved in the user specified directory, crun#, which is located inside the project directory. These files include chemical shifts (belok.prot file), sequence file, peak lists, dihedral angles constraints (file belok.aco), H-bond constraints, if available, (file hbond.upl), and constraints for ZN ions, if present, (files zn.upl, zn.lol).
Step 5. Structure evaluation and peak lists refinement
RPF analysis and DP score allow one to estimate goodness-of-fit of a structural ensemble to NOESY peak lists. The results of RPF analysis can serve both for structure validation and peak lists refinement.
- run RPF analysis { Structure>RPF>RP }. The results of the RPF analysis include peak lists in the SPARKY format of both false negative and false positive peaks for C13-aliphatic NOESY, C13-Aromatic NOESY, and N15-NOESY spectra in separate files.
- set up DP-score calculations with AutoStructure { Structure>RPF>DP }.