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By Vikram K. Mulligan, Baker laboratory. Documentation written 4 April 2014, and last updated on 11 March 2016.

Short summary

GeneralizedKIC (short for Generalized Kinematic Closure) is a generalization of the existing kinematic closure machinery. The generalized version works with arbitrary backbones, and with loops that go through side-chains, ligands, etc. It has also been implemented with full RosettaScripts support, making it easy to incorporate into more complicated protocols. The RosettaScripts interface gives full control over the inner workings of the algorithm, making it quite a powerful and versatile tool.

Usage cases

Generalized KIC is useful for the following situations:

  • Given an arbitrary unbranched, covalently-connected part of a structure, with well-defined, fixed starting and ending points, one wishes to sample alternative conformations that preserve ideal geometry. An example would be a case in which the N- and C-termini of a protein were linked by a disulfide bond, and one wished to sample alternative conformations for a short stretch of the N-terminal backbone, the disulfide bond, and a short stretch of the C-terminal backbone, subject to the condition that the disulfide remains closed.
  • Given a part of a structure that is open but which ought to be covalently connected, with well-defined, fixed starting and ending points, one wishes to sample conformations that close the segment. For example, given unjoined antiparallel beta strands forming a two-strand sheet, one might wish to sample hairpin conformations for a loop connecting the strands. This closure can be through side-chains or ligands, too: given two segments containing metal-binding residues, one might want to sample conformations that allow a metal to be bound with ideal geometry.
  • Given a covalent linkage between distant parts of a FoldTree and a structural perturbation that pulls the atoms involved in the covalent linkage apart, one wishes to find compensatory structural perturbations that maintain the closed covalent geometry while minimally altering the structure.

The above scenarios tend to be sub-problems of more complicated problems, particularly involving heavily cross-linked molecules for which one might wish to sample many conformations.

Input and output

GeneralizedKIC takes as input a pose that contains a covalently-connected chain of residues (a "loop", where the loop need not be connected solely by conventional backbone connections) to which the algorithm will be applied. The chain of residues need not start out in a sensible conformation, nor need it consist of canonical alpha-amino acids.

GeneralizedKIC will return a pose in which the loop in question has been put in a new, closed conformation (subject to user-specified sampling methods). Only the loop residues (and any "tail" residues specified) will be moved by this mover; all other residues will remain in their starting positions. The FoldTree is disregarded by the GeneralizedKIC mover, and will not be altered by it. (Note: if a pre-selection mover is defined as described in the GeneralizedKIC selector documentation, this mover can alter geometry outside of the loop to be closed, or could alter the FoldTree.)

If no closed solution could be found, GeneralizedKIC will return the original input pose. If a ContingentFilter has been supplied, its value will be set to "false" in this case (and "true" otherwise). By default, GeneralizedKIC will set the mover status to failure if no solution is found, but the user can override this with dont_fail_if_no_solution_found=true.

General workflow

In general, one must:

  1. Build or import a structure, which must include the loop residues to be closed (though these need not be in a closed conformation to start).
  2. Ensure that covalent linkages have been declared with the DeclareBond mover.
  3. Set the GeneralizedKIC options.
  4. Define the loop to be closed, and set pivots.
  5. Set up one or more GeneralizedKIC perturbers, which determine how loop conformations will be sampled.
  6. Define one or more GeneralizedKIC filters.
  7. Set the GeneralizedKIC selector used to choose a solution from among those found (and an optional pre-selection mover).

These steps are discussed in detail in the next section.

Detailed workflow

  1. Build or import a structure. GeneralizedKIC cannot add residues or geometry. If one wants to build a new loop, for example, one must add the new loop residues before calling GeneralizedKIC. (In RosettaScripts, the PeptideStubMover is useful for building geometry from scratch or for adding residues to existing geometry.)

  2. Ensure that covalent linkages have been declared. GeneralizedKIC will move atoms about to ensure ideal geometry, but cannot declare new chemical bonds. (In RosettaScripts, the DeclareBond mover lets Rosetta know that certain residues are covalently attached to one another.)

  3. Set the GeneralizedKIC options (number of closure attempts, whether the algorithm should accept the first successful closure or choose from all successful closure attempts, etc.). In RosettaScripts, this is handled inside the block as follows:

    <MOVERS>
    ...
         <GeneralizedKIC name="&string" low_memory_mode="(false &bool)" closure_attempts="(100 &int)" stop_if_no_solution="(0 &int)" stop_when_n_solutions_found="(0 &int)" selector="&string" selector_scorefunction="&string" selector_kbt="(1.0 &Real)" contingent_filter="&string" dont_fail_if_no_solution_found="(false &bool)" correct_polymer_dependent_atoms="(false &bool)">
              ...
         </GeneralizedKIC>
    ...
    </MOVERS>
    The low_memory_mode option can be used to limit the amount of information about each solution found that is stored, in order to reduce memory consumption. See the note below for more details about the advantages and risks of this non-default mode. The closure_attempts parameter sets the number of times the algorithm will try to close the loop. A setting of 0 means that it will keep trying indefinitely. The stop_when_n_solutions_found option allows the algorithm to stop after finding at least N successful solutions, or, if this is set to 0, to keep going until it has done as many attempts as specified by closure_attempts; in either case a solution is then chosen by the selector. (Note that, because a single attempt returns up to 16 closure solutions, the selector will be applied even if stop_when_n_solutions_found is set to 1, since more than one solution might have been found in the first successful attempt). The selector flag is mandatory, and specifies the way in which a solution is chosen from among the successful solutions. The selector_scorefunction flag allows a separate scorefunction to be used by those selectors that select based on energy or score; this is recommended since score terms based on side-chain packing may produce poor results, since the GeneralizedKIC algorithm does not call the packer. Some selectors also take a temperature value, set by the selector_kbt option. See the GeneralizedKIC selector documentation for more details.
    In some cases, the GeneralizedKIC mover will find no solution. This could be because no solution exists (e.g. if the loop is too short for the endpoint separation, or if there is geometry blocking any path between the endpoints), because the sampling method used was too restrictive, or because too few attempts were made. If this happens, the pose is left unaltered. If the loop geometry is open, it is useful to have a means of aborting the trajectory in this case. By default, the mover returns failure status, aborting the trajectory. This can be overridden by setting dont_fail_if_no_solution_found=true. A ContingentFilter can also be used to record whether the mover failed. The ContingentFilter is a specialized filter that has its value set by a mover. GeneralizedKIC can set the value of a ContingentFilter, specified using the contingent_filter flag, to true or false depending on whether the closure was successful or unsuccessful. Subsequent application of the filter, possibly at a later point, can then abort trajectories involving unsuccessful loop closure. As a final note, GeneralizedKIC can correct positions of atoms that depend on polymer bonds (such as amide protons, carbonyl oxygens, or N-methyl groups in N-methylated amino acids) to ideal positions if the correct_polymer_dependent_atoms option is set to "true".
  4. Define a series of residues for the GeneralizedKIC closure problem. This must be an unbranched chain of residues with continuous covalent linkages, listed in order from one end of the chain to the other. When the GeneralizedKIC::apply() function is called, a continuous chain of atoms running through the selected residues is automatically chosen. Residues are specified with AddResidue tags within a GeneralizedKIC block. Pivot points must also be indicated explicitly, using the SetPivots tag. Pivots are atoms in the chain of atoms to be closed that are flanked by bonds whose dihedral values will be solved for analytically by the closure algorithm in order to close the loop. Currently, due to hard-coded assumptions in the kinematic closure numerical library, the first pivot must be the second atom in the chain to be closed, and the last pivot must be the second-to-last atom in the chain to be closed. This restriction will be eliminated in a future version of GeneralizedKIC.
    Optionally, additional "tail" residues can also be listed. These are residues that are either connected directly to the loop to be closed, or connected indirectly to this loop through other tail residues, and which move with the loop to be closed. The AddTailResidue tag can be used to specify these. Order is not important for the AddTailResidue flag.

    <MOVERS>
    ...
         <GeneralizedKIC ...>
              <AddResidue res_index="(&int)" />
              <AddResidue res_index="(&int)" />
              <AddResidue res_index="(&int)" />
              ...
              <AddTailResidue res_index="(&int)" />
              <AddTailResidue res_index="(&int)" />
              <AddTailResidue res_index="(&int)" />
              ...
              <SetPivots res1="(&int)" atom1="&string" res2="(&int)" atom2="&string" res3="(&int)" atom3="&string" />
              ...
         </GeneralizedKIC>
    ...
    </MOVERS>
    For example, if one were closing a loop consisting of residues ALA44, CYS45, LYS46, CYS47, CYS23, ASP22, and PHE21, where CYS47 and CYS23 were linked by a disulfide bond, and where CYS45 were linked to CYS12 by another disulfide such that CYS12 was expected to move with CYS45, one would write:
    <MOVERS>
    ...
         <GeneralizedKIC ...>
              <AddResidue res_index="4" />
              <AddResidue res_index="45" />
              <AddResidue res_index="46" />
              <AddResidue res_index="47" />
              <AddResidue res_index="23" />
              <AddResidue res_index="22" />
              <AddResidue res_index="21" />
              <AddTailResidue res_index="12" />
              <SetPivots res1="4" atom1="CA" res2="23" atom2="SG" res3="21" atom3="CA" />
              ...
         </GeneralizedKIC>
    ...
    </MOVERS>
    From the above example, we can see that loop segments may run backwards or forwards, or may involve residues that are far apart in linear sequence provided they are covalently linked. Note that while the sequence of residues matter, the overall direction of the loop does not: we could just as happily have added residues in the reverse order (21->22->23->47->46->45->44). In this example, we have arbitrarily chosen CYS23's SG atom as the middle pivot point, though any atom in the chain that is flanked by bonds that can rotate freely could have been chosen.
  5. Define one or more GeneralizedKIC perturbers. Each perturber samples conformation space for each closure attempt. See GeneralizedKIC perturbers for details.

  6. Define one or more GeneralizedKIC filters. Filters are applied after each closure attempt, and eliminate solutions that don't meet some criterion. See GeneralizedKIC filters for details.

  7. Set the GeneralizedKICselector. The selector chooses a single solution from the set of solutions found that pass all of the filters. See the documentation on GeneralizedKIC selectors for details, including details on the optional pre-selection mover (a mover applied to all solutions passing filters prior to application of the selector).

That's it! You should be happily closing loops, now.

Algorithm details

The kinematic closure algorithm is described in detail in Coutsias et al. (2004). J. Comput. Chem. 25(4):510-28. and in Mandell et al. (2009). Nat. Methods. 6(8)551-2.. Briefly, given a rigid-body transform describing the position and orientation of the residue immediately following a loop relative to a residue immediately preceding a loop, a loop with N degrees of freedom now has N-6 effective degrees of freedom, the remaining 6 degrees of freedom being determined by the condition that the loop be closed. This means that all but 6 degrees of freedom of the loop (e.g. all but 6 chain dihedral angles) may be sampled, perturbed, or set as one sees fit, and the remaining 6 degrees of freedom may be solved for analytically to impose the closure condition. The GeneralizedKIC algorithm does just this, allowing the user to sample all but 6 degrees of freedom of the loop, then solve analytically for the remainining 6 (the dihedral angles flanking 3 pivot atoms) to ensure that the loop is closed. For any given set of values for the other N-6 degrees of freedom, there may be anywhere from 0 to 16 solutions to the system of equations solved by the kinematic closure algorithm.

Low-memory mode

By default, all solutions found by GenKIC are stored as full poses until one is selected by the selector. Unfortunately, this means that if instructed to store many solutions, GenKIC can exhaust available memory. As of 13 March 2016, a low-memory mode has been added ("low_memory_mode=true"). In this mode, only loop degree-of-freedom values are stored prior to selection. The drawback, however, is it can be risk to use low-memory mode with a preselection mover. Any changes to the pose made by the preselection mover will not be stored, so the preselection mover will have to be re-applied to the selected pose after selection. This costs additional time. In addition, if the preselection mover has any stochastic component to its behaviour, then the second application may not produce identical results to the first. This means that what might have been the lowest-energy pose when the preselection mover was first applied could now be a relatively high-energy pose, for example. For this reason, be judicious in the use of low-memory mode. The preferred course is to stop GenKIC when N solutions have been found to limit memory usage.

Example RosettaScripts

This example creates a 10-residue cyclic peptide with a disulfide bond between the N- and C-termini. It then defines a loop starting at residue 3, going backwards to residue 1, through the disulfide to residue 10, and back to residue 5. This loop is closed by kinematic closure, with loop conformations sampled. A bump check filter is applied (AddFilter type="loop_bump_check"), and a solution chosen randomly (selector="random_selector") from the solutions found.

<ROSETTASCRIPTS>
    <TASKOPERATIONS>
    </TASKOPERATIONS>
    <SCOREFXNS>
        <ScoreFunction name="tala" weights="talaris2013" symmetric="0" />
    </SCOREFXNS>
    <FILTERS>
    </FILTERS>
    <MOVERS>
        <PeptideStubMover name="pep_stub" reset="1">
            <Append resname="CYD" />
            <Append resname="ALA" />
            <Append resname="ALA" />
            <Append resname="ALA" />
            <Append resname="ALA" />
            <Append resname="ALA" />
            <Append resname="ALA" />
            <Append resname="ALA" />
            <Append resname="ALA" />
            <Append resname="CYD" />
        </PeptideStubMover>
        
        <SetTorsion name="tor1">
            <Torsion residue="ALL" torsion_name="phi" angle="-64.7"/>
            <Torsion residue="ALL" torsion_name="psi" angle="-41.0"/>
            <Torsion residue="ALL" torsion_name="omega" angle="180"/>
        </SetTorsion>

	<SetTorsion name="tor2">
            <Torsion residue="pick_atoms" angle="random">
                <Atom1 residue="1" atom="N"/>
                <Atom2 residue="1" atom="CA"/>
                <Atom3 residue="1" atom="CB"/>
                <Atom4 residue="1" atom="SG"/>
            </Torsion>
            <Torsion residue="pick_atoms" angle="random">
                <Atom1 residue="10" atom="N"/>
                <Atom2 residue="10" atom="CA"/>
                <Atom3 residue="10" atom="CB"/>
                <Atom4 residue="10" atom="SG"/>
            </Torsion>
	</SetTorsion>
        
        <DeclareBond name="bond" res1="1" atom1="SG" res2="10" atom2="SG"/>

	<GeneralizedKIC name="genkic" closure_attempts="200" stop_when_n_solutions_found="0" selector="random_selector">
		<AddResidue res_index="3" />
                <AddResidue res_index="2" />
                <AddResidue res_index="1" />
                <AddResidue res_index="10" />
                <AddResidue res_index="9" />
                <AddResidue res_index="8" />
                <AddResidue res_index="7" />
                <AddResidue res_index="6" />
                <AddResidue res_index="5" />
		<SetPivots res1="3" atom1="CA" res2="1" atom2="SG" res3="5" atom3="CA" />
		<CloseBond prioratom_res="10" prioratom="CB" res1="10" atom1="SG" res2="1" atom2="SG" followingatom_res="1" followingatom="CB" bondlength="2.05" angle1="103" angle2="103" randomize_flanking_torsions="true" />
		<AddPerturber effect="randomize_alpha_backbone_by_rama">
			<AddResidue index="3" />
			<AddResidue index="2" />
			<AddResidue index="1" />
			<AddResidue index="10" />
			<AddResidue index="9" />
                        <AddResidue index="8" />
                        <AddResidue index="7" />
                        <AddResidue index="6" />
                        <AddResidue index="5" />
		</AddPerturber>
		<AddFilter type="loop_bump_check" />
	</GeneralizedKIC>
        
    </MOVERS>
    <APPLY_TO_POSE>
    </APPLY_TO_POSE>
    <PROTOCOLS>
        <Add mover="pep_stub"/>
        <Add mover="tor1"/>
        <Add mover="tor2"/>
        <Add mover="bond"/>
        <Add mover="genkic"/>
    </PROTOCOLS>
</ROSETTASCRIPTS>

Failure cases

GeneralizedKIC cannot handle the following cases:

  • Multiple covalent connections between a single pair of residues in the chain of residues to be closed. (This is something that, in general, Rosetta handles poorly).
  • Loops involving salt bridges, cation-pi interactions, hydrogen bonds, or other noncovalent interactions. (The GeneralizedKIC framework has been written with this in mind as a possible future extension, however.) Note that coordinate covalent bonds between metal-binding residues and metal ions are considered "covalent" in Rosetta, and are handled properly by GeneralizedKIC.

Full options list

Autogenerated Tag Syntax Documentation:


The GeneralizedKIC mover allows a user to define a chain of atoms whose conformation is to be sampled or altered subject to the constraint that it remain closed, or that it be closed if it starts out open. The mover has myriad options for controlling sampling and selection of a solution, and can invoke other movers to apply to each closed solution that it finds. It has been written to be fully general, and can operate on chains of atoms that pass through backbones, side-chains, ligands, etc. No assumptions about alpha-amino acid backbones are hard-coded, so GeneralizedKIC should be fully compatible with any non-canonical entity.

<GeneralizedKIC name="(&string;)" low_memory_mode="(&bool;)"
        correct_polymer_dependent_atoms="(&bool;)"
        dont_fail_if_no_solution_found="(&bool;)"
        selector="(&genkic_selector_name;)" selector_scorefunction="(&string;)"
        selector_kbt="(&real;)" stop_if_no_solution="(0 &non_negative_integer;)"
        closure_attempts="(&non_negative_integer;)"
        stop_when_n_solutions_found="(&non_negative_integer;)"
        pre_selection_mover="(&string;)" contingent_filter="(&string;)" >
    <AddResidue res_index="(&non_negative_integer;)" />
    <AddTailResidue res_index="(&non_negative_integer;)" />
    <SetPivots res1="(&non_negative_integer;)" atom1="(&string;)"
            res2="(&non_negative_integer;)" atom2="(&string;)"
            res3="(&non_negative_integer;)" atom3="(&string;)" />
    <SampleCisPeptideBond cis_prob="(0.1 &real;)" >
        <AddResidue index="(&non_negative_integer;)" />
    </SampleCisPeptideBond>
    <CloseBond res1="(&non_negative_integer;)" atom1="(&string;)"
            res2="(&non_negative_integer;)" atom2="(&string;)" bondlength="(&real;)"
            angle1="(&real;)" angle2="(&real;)"
            randomize_flanking_torsions="(&bool;)"
            prioratom_res="(&non_negative_integer;)" prioratom="(&string;)"
            followingatom_res="(&non_negative_integer;)" followingatom="(&string;)"
            torsion="(&real;)" />
    <AddPerturber effect="(&genkic_perturber_name;)"
            bin_params_file="(ABBA &string;)"
            iterations="(1 &non_negative_integer;)"
            must_switch_bins="(false &bool;)" bin="(&string;)"
            custom_rama_table="(&string;)" >
        <AddResidue index="(&non_negative_integer;)" />
        <AddAtoms res1="(&non_negative_integer;)" atom1="(&string;)"
                res2="(&non_negative_integer;)" atom2="(&string;)"
                res3="(&non_negative_integer;)" atom3="(&string;)"
                res4="(&non_negative_integer;)" atom4="(&string;)" />
        <AddValue value="(&real;)" />
    </AddPerturber>
    <AddFilter type="(&genkic_filter_name;)" residue="(&non_negative_integer;)"
            bin_params_file="(ABBA &string;)" bin="(&string;)"
            rama_cutoff_energy="(0.3 &real;)" >
        <AddFilterParameterReal name="(&string;)" value="(&real;)" />
        <AddFilterParameterInteger name="(&string;)" value="(&non_negative_integer;)" />
        <AddFilterParameterBoolean name="(&string;)" value="(&bool;)" />
        <AddFilterParameterString name="(&string;)" value="(&string;)" />
    </AddFilter>
    <AddAtomPairDistanceFilter res1="(&non_negative_integer;)" atom1="(&string;)"
            res2="(&non_negative_integer;)" atom2="(&string;)" distance="(&real;)"
            greater_than="(&bool;)" />
</GeneralizedKIC>
  • low_memory_mode: The low_memory_mode option can be used to limit the amount of information about each solution found that is stored, in order to reduce memory consumption. In this mode, only loop degree-of-freedom values are stored prior to selection. The drawback, however, is it can be risk to use low-memory mode with a preselection mover. Any changes to the pose made by the preselection mover will not be stored, so the preselection mover will have to be re-applied to the selected pose after selection. This costs additional time. In addition, if the preselection mover has any stochastic component to its behaviour, then the second application may not produce identical results to the first. This means that what might have been the lowest-energy pose when the preselection mover was first applied could now be a relatively high-energy pose, for example. For this reason, be judicious in the use of low-memory mode. The preferred course is to stop GenKIC when N solutions have been found to limit memory usage.
  • correct_polymer_dependent_atoms: If set to true, atoms whose positions are dependent on polymer bonds, such as amino acid "H" and "O" atoms, will have their positions set to ideal coordinates after closure. False by default.
  • dont_fail_if_no_solution_found: By default, the GeneralizedKIC mover returns failure status if it fails to find a closed solution that passes filters. Certain usage cases require that it does not fail. If this option is set to true, it prevents GeneralizedKIC from returning a failure status, even if it finds no solution (in which case the input pose, which may have an open chain of atoms, is returned as the output pose). This is necessary, for example, when GeneralizedKIC is used in conjunction with a ContingentFilter.
  • selector: (REQUIRED) The GeneralizedKICselector to use to pick a solution. Solutions may be picked randomly ("random_selector"), by energy ("lowest_energy_selector"), or by other criteria. See the Rosetta help wiki for more information on GeneralizedKICselectors, and for the full range of options.
  • selector_scorefunction: A scorefunction that the GeneralizedKICselector will use to select the solution to return.
  • selector_kbt: If the GeneralizedKICselector is set to "boltzmann_energy_selector", this is the Boltzmann temperature used when selecting a solution randomly, weighted by the Boltzmann probability of that solution.
  • stop_if_no_solution: If this option is used and set to a positive integer N, GeneralizedKIC stops looking for closure solutions if no solution was found in the first N attempts.
  • closure_attempts: The maximum number of times to try to find a solution. Set this to zero to keep trying indefinitely (in which case the "stop_when_n_solutions_found" option should be used to prevent the mover from iterating forever).
  • stop_when_n_solutions_found: If this option is used and set to a positive integer N, GeneralizedKIC will stop looking for closure solutions as soon as at least N solutions have been found that pass filters. Note that a GeneralizedKICselector must be specified even if this option is set to 1, because a single attempt can yield up to 16 solutions.
  • pre_selection_mover: As an optional alternative to restricting oneself to backbone-only score terms, GeneralizedKIC permits the user to specify a pre-selection mover defined prior to the GeneralizedKIC block. This permits side-chain moves, such as repacking or side-chain minimization, prior to scoring with the full energy function. Note that this mover will be applied to all candidate solutions passing filters, which makes this option potentially very computationally expensive! If a pre-selection mover is specified, the mover may alter geometry outside of the loop to be closed, or may alter the FoldTree, in which case these alterations will carry through to the final pose returned by GeneralizedKIC. TaskOperations and MoveMaps may be used appropriately in the definition of the mover in question to restrict its effects to the loop on which GeneralizedKIC is operating if the user does not wish to alter geometry outside of this loop. If one wishes to apply more than one pre-selection mover (for example, a PackRotamersMover followed by a sidechain-only MinMover, to repack and minimize side-chains), they may be combined in a ParsedProtocol mover. As a final note, a mover may return failure status, or may encapsulate filters that return failure status, in which case the solution to which the mover has been applied will be discarded.
  • contingent_filter: A ContingentFilter can also be used to record whether the mover failed. The ContingentFilter is a specialized filter that has its value set by a mover. GeneralizedKIC can set the value of a ContingentFilter, specified using the contingent_filter flag, to true or false depending on whether the closure was successful or unsuccessful. Subsequent application of the filter, possibly at a later point, can then abort trajectories involving unsuccessful loop closure. Note that this is a largely deprecated feature that has been retained since a few users have found it useful in some cases; for most usage cases, one can simply rely on the fact that the mover returns failure status if no closed solution is found. It will likely be necessary to use the "dont_fail_if_no_solution_found" option if a ContingentFilter is used.

Subtag AddResidue: "AddResidue" sub-tags are used to specify the residues that make up the chain of atoms to be closed, in order of connectivity.

  • res_index: (REQUIRED) The "res_index" option within an "AddResidue" sub-tag specifies the index, in Rosetta numbering, of the residue being added to the chain of residues to be closed.

Subtag AddTailResidue: Loops to be closed might have other chains of residues attached to them. By default, GeneralizedKIC fails to respect covalent bonds to any but the end residues in the chain to be closed, and can tear these bonds apart as residues move. Users can optionally specify a list of "tail" residues that are covalently attached to, and move with, the chain of residues to be closed. For example, one might have a series of glycans attached to a serine residue in a protein loop that's being closed with GenKIC. If these residues are specified as tail residues, then they will remain attached to the serine residue even as the loop that contains the serine residue moves.

  • res_index: (REQUIRED) The "res_index" option within an "AddTailResidue" sub-tag specifies the index, in Rosetta numbering, of the residue being added to the chain of residues constituting a "tail" that is intended to move with the chain to be closed.

Subtag SetPivots: Pivots are atoms in the chain of atoms to be closed that are flanked by bonds whose dihedral values will be solved for analytically by the closure algorithm in order to close the loop. Currently, due to hard-coded assumptions in the kinematic closure numerical library, the first pivot must be the second atom in the chain to be closed, and the last pivot must be the second-to-last atom in the chain to be closed.

  • res1: (REQUIRED) The first pivot residue (by Rosetta index). Currently, due to hard-coded assumptions in the kinematic closure numerical library, the first pivot must be the second atom in the chain to be closed.
  • atom1: (REQUIRED) The first pivot atom (by atom name). Currently, due to hard-coded assumptions in the kinematic closure numerical library, the first pivot must be the second atom in the chain to be closed.
  • res2: (REQUIRED) The middle pivot residue (by Rosetta index).
  • atom2: (REQUIRED) The middle pivot atom (by atom name).
  • res3: (REQUIRED) The last pivot residue (by Rosetta index). Currently, due to hard-coded assumptions in the kinematic closure numerical library, the last pivot must be the second-to-last atom in the chain to be closed.
  • atom3: (REQUIRED) The last pivot atom (by atom name). Currently, due to hard-coded assumptions in the kinematic closure numerical library, the last pivot must be the second-to-last atom in the chain to be closed.

Subtag SampleCisPeptideBond: A "SampleCisPeptideBond" sub-tag is a shorthand for adding a sample_cis_peptide_bond GeneralizedKICperturber. It tells the GeneralizedKIC mover to sample a cis-peptide bond at a particular residue some subset of the time.

  • cis_prob: The probability of sampling a cis-peptide bond at a given position, where 1.0 means that cis is sampled 100% of the time, and 0.0 means that trans is sampled 100% of the time.

Subtag AddResidue: "AddResidue" sub-sub-tags are used within a "SampleCisPeptideBond" sub-tag to list all of the residues at which cis-peptide bonds should be sampled.

  • index: (REQUIRED) The index, in Rosetta numbering, of a residue whose omega value should sample 0.

Subtag CloseBond: A "CloseBond" sub-tag is a shorthand for set_bondangle and set_bondlength GeneralizedKICperturbers, as well as an optional pair of randomize_dihedral perturbers for the flanking dihedral angles, and an optional set_torsion perturber. It is intended to be used to set the ideal geometry for an open bond in a loop that GeneralizedKIC should close.

  • res1: (REQUIRED) The residue (Rosetta numbering) containing the first atom in the bond to be closed.
  • atom1: (REQUIRED) The first atom (given as an atom name string) in the bond to be closed.
  • res2: (REQUIRED) The residue (Rosetta numbering) containing the second atom in the bond to be closed.
  • atom2: (REQUIRED) The second atom (given as an atom name string) in the bond to be closed.
  • bondlength: (REQUIRED) The length of the bond to be closed.
  • angle1: (REQUIRED) The bond angle defined by B1, A1, A2, where A1 and A2 are the atoms in the bond to be closed and B1 is the last atom in the chain to be closed that's prior to the bond to be closed.
  • angle2: (REQUIRED) The bond angle defined by A1, A2, B2, where A1 and A2 are the atoms in the bond to be closed and B2 is the first atom in the chain to be closed that's past the bond to be closed.
  • randomize_flanking_torsions: If true, bonds flanking the bond to be closed will have their torsion values randomized. If present, requires "prioratom_res" and "followingatom_res" attributes to be set.
  • prioratom_res: The atom preceding the bond to be closed. This is only required if the "randomize_flanking_torsions" option is used.
  • prioratom: The atom preceding the bond to be closed. This is only required if the "randomize_flanking_torsions" option is used.
  • followingatom_res: The atom following the bond to be closed. This is only required if the "randomize_flanking_torsions" option is used.
  • followingatom: The atom following the bond to be closed. This is only required if the "randomize_flanking_torsions" option is used.
  • torsion: An optional torsion value for the bond to be closed. If not specified, the input value persists.

Subtag AddPerturber: The "AddPerturber" sub-tag is used to add and configure GeneralizedKICperturbers. GeneralizedKICperturbers alter the chain to be closed in some way prior to kinematic closure, allowing definition of a desired conformation or conformational sampling. They can only act on the chain to be closed, and have no effect on tail residues or on any other part of the input structure. Perturbers are applied in the order that they are defined. Different perturbers may alter the same degrees of freedom, sequentially.

  • effect: (REQUIRED) The name of the GeneralizedKICperturber (e.g. "perturb_dihedral", "set_bondangle", "randomize_alpha_backbone_by_rama", etc.). See the Rosetta documentation wiki for a full list of available perturbers.
  • bin_params_file: The "randomize_backbone_by_bins" and "perturb_backbone_by_bins" perturbers require a bin definition file, set using this option.
  • iterations: The "perturb_backbone_by_bins" perturber requires that a number of iterations be set. If the number of iterations is set to 1, only one residue, randomly chosen from the list provided, will have its mainchain torsions perturbed. If it is set higher, the algorithm iteratively picks a residue at random and perturbs it.
  • must_switch_bins: This option is only used by the "perturb_backbone_by_bins" perturber. If "must_switch_bins" is set to true, the chosen residue is forced into a different torsion bin; if false, it has some probability of remaining in the same torson bin, in which case its mainchain torsion values will be chosen randomly (in a biased manner, if possible) from within that bin.
  • bin: The "set_backbone_bin" perturber draws random mainchain torsion values from a mainchain torsion bin defined in a bin params file. The "bin" option names the bin from which the torsion values will be drawn. Required for this perturber only.
  • custom_rama_table: The "randomize_alpha_backbone_by_rama" perturber may optionally draw random phi and psi values from one of several custom Ramachandran tables that offer either flattened or more stringent distributions. These are listed on the Rosetta help wiki.

Subtag AddResidue: The "AddResidue" sub-sub-tag in an "AddPerturber" sub-tag permits a residue to be defined on which the perturber may act. Use multiple "AddResidue" tags to define more than one residue.

  • index: (REQUIRED) The index, in Rosetta numbering, of a residue on which this perturber should act.

Subtag AddAtoms: The "AddAtoms" sub-sub-tag in an "AddPerturber" sub-tag permits one or more atoms to be defined, on which the perturber may act. A particular type of perturber may take a list of one, two, three, or four atoms, depending on what it acts on.

  • res1: (REQUIRED) The residue (specified using Rosetta numbering) containing the first atom.
  • atom1: (REQUIRED) The first atom, specified as an atom name string.
  • res2: The residue (specified using Rosetta numbering) containing the second atom.
  • atom2: The second atom, specified as an atom name string.
  • res3: The residue (specified using Rosetta numbering) containing the third atom.
  • atom3: The third atom, specified as an atom name string.
  • res4: The residue (specified using Rosetta numbering) containing the fourth atom.
  • atom4: The fourth atom, specified as an atom name string.

Subtag AddValue: The "AddValue" sub-sub-tag in an "AddPerturber" sub-tag permits a value to be set for those perturbers that take a value. (An example of this is the "set_dihedral" perturber, which takes a value for the dihedral angle to set.) Use multiple "AddValue" tags to specify more than one value.

  • value: (REQUIRED) The value to pass to the perturber.

Subtag AddFilter: An "AddFilter" sub-tag adds a GeneralizedKICfilter, which discards solutions that do not pass certain criteria. Generally, GeneralizedKICfilters are written to be considerably faster than conventional Rosetta filters, since they usually do not act on full poses. They are applied before pre-selection movers.

  • type: (REQUIRED) The name of the GeneralizedKICfilter to apply (e.g. "backbone_bin", "rama_prepro_check", "atom_pair_distance", etc.). See the Rosetta help wiki for a full list of avaliable filters.
  • residue: A parameter required by several filters, including "backbone_bin", "alpha_aa_rama_check", and "rama_prepro_check": the index, in Rosetta numbering, of the residue that we're examining with this filter.
  • bin_params_file: A parameter specific for the "backbone_bin" filter: a filename for a file defining mainchain torsion bins.
  • bin: A parameter specific for the "backbone_bin" filter: the mainchain torsion bin in which an amino acid residue must lie. If the residue is not in this bin, the solution is discarded.
  • rama_cutoff_energy: A parameter specific for the "alpha_aa_rama_check" and "rama_prepro_check" filters: the value for the rama or rama_prepro score terms above which solutions are discarded.

Subtag AddFilterParameterReal: An "AddFilterParameterReal" sub-sub-tag in an "AddFilter" tag sets a real-valued setting for a filter. For example, the "atom_pair_distance" filter takes a real-valued parameter, called "distance", for the cutoff separation between two atoms used to discard solutions.

  • value: (REQUIRED) The value of the real-valued parameter.

Subtag AddFilterParameterInteger: An "AddFilterParameterInteger" sub-sub-tag in an "AddFilter" tag sets an integer setting for a filter. For example, the "atom_pair_distance" filter takes two integer parameters, called "res1" and "res2", for the indices of the two residues containing the two atoms whose separation is to be measured.

  • value: (REQUIRED) The value of the integer parameter.

Subtag AddFilterParameterBoolean: An "AddFilterParameterBoolean" sub-sub-tag in an "AddFilter" tag sets a Boolean setting for a filter. For example, the "atom_pair_distance" filter has a Boolean parameter called "greater_than" which, when set to "true" selects for interatomic separation greater than a cutoff rather than less than the cutoff.

  • value: (REQUIRED) The value ("true" or "false") of the Boolean parameter.

Subtag AddFilterParameterString: An "AddFilterParameterString" sub-sub-tag in an "AddFilter" tag sets a string setting for a filter. For example, the "atom_pair_distance" filter takes two string parameters, called "atom1" and "atom2", for the names of the two atoms whose separation is to be measured.

  • value: (REQUIRED) The value of the string parameter.

Subtag AddAtomPairDistanceFilter: The "AddAtomPairDistanceFilter" sub-tag is a shorthand for adding an "atom_pair_distance" GeneralizedKICFilter. It discards solutions if pairs of atoms are separated by more than a cutoff distance.

  • res1: (REQUIRED) The residue (Rosetta index) containing the first atom in the pair whose separation we will be measuring.
  • atom1: (REQUIRED) The first atom in the pair whose separation we will be measuring, specified as an atom name string.
  • res2: (REQUIRED) The residue (Rosetta index) containing the second atom in the pair whose separation we will be measuring.
  • atom2: (REQUIRED) The second atom in the pair whose separation we will be measuring, specified as an atom name string.
  • distance: (REQUIRED) The cutoff inter-atomic distance for discarding solutions.
  • greater_than: If "greater_than" is set to true, the filter will discard any solution for which the atoms are not separated by AT LEAST the cutoff distance. If it is false (the default), then any solution for which the atoms are separated by more than the cutoff distance is discarded.