Using residue patches in AbinitioRelax

I am hoping to ‘fold’ short phosphorylated peptide sequences, and started by generating fragments with SS predictions on the unphosphorylated peptides. For example, I ran this sequence through a local installation of PSIPRED and picked 200 3- and 9-mers:

>1

MKGDAHRYLAEFATG

I checked that the TYR_p:phosphorylated patch was active in the full-atom patches.txt, then created a centroid patch and added it to the patches.txt by analogy to existing patches:

NAME phosphorylated

TYPES PHOSPHORYLATION

BEGIN_SELECTOR

PROPERTY PROTEIN

AA TYR

NOT VARIANT_TYPE PHOSPHORYLATION ## Don’t phosphorylate a residue twice

END_SELECTOR

BEGIN_CASE ### THE GENERAL CASE ##########################################

ADD_PROPERTY CHARGED ## For the phosphate group

END_CASE

If I modify the centroid patch, I get an error that a residue type cannot be found for TYR_p:phosphorylated, so it is definitely doing something. 

I have then been attempting to run AbinitioRelax on the phosphorylated fasta:

>1

MKGDAHR[TYR_p:phosphorylated]LAEFATG

And get weird output and errors:

protocols.abinitio.AbrelaxApplication: read fasta sequence: 36 residues

MKGDAHR[TYR_p:phosphorylated]LAEFATG

…

core.chemical.Patch: Reading patch file: /export/ActiveUsers/kentgorday/rosetta_src_2018.33.60351_bundle/main/database/chemical/residue_type_sets/centroid/patches/tyr_phosphorylated.txt

…

core.pose: one_letter: MKGDAHRLAEFATG

core.pose: seq_in: MKGDAHR[TYR_p:phosphorylated]LAEFATG

core.chemical.ResidueTypeFinder: No ResidueTypes remain after filtering for matched variants.

core.chemical.ResidueTypeFinder: No ResidueTypes remain after filtering for matched variants.

…

core.pose: residue_types_from_sequence():  seqpos: 6 aa H HIS

core.chemical.PatchOperations.hh: AddProperty::apply: CHARGED

core.chemical.Patch: successfully patched: TYR to: TYR:phosphorylated

core.pose: residue_types_from_sequence():  seqpos: 7 aa R ARG

…

core.pose: make_pose_from_sequence():  seqpos: 6 HIS

core.conformation.Conformation: CURRENT_FOLD_TREE  EDGE 1 6 -1

core.pose: make_pose_from_sequence():  seqpos: 7 TYR

core.conformation.Conformation: CURRENT_FOLD_TREE  EDGE 1 7 -1

core.pose: make_pose_from_sequence():  seqpos: 8 LEU

core.conformation.Conformation: CURRENT_FOLD_TREE  EDGE 1 8 -1

…

protocols.abinitio:

===================================================================

   Stage 1

   Folding with score0 for max of 20000

protocols.simple_moves.FragmentMover:  1 2 3 4 5 6 7size of insertmap: 7 — insert_size:  9 9 9 9 9 9 9

protocols.simple_moves.FragmentMover: [ ERROR ] BEGIN: 7 SIZE: 9 TOTAL_RES: 14

protocols.simple_moves.FragmentMover: [ ERROR ] Are the fragments compatible with the fasta or the input PDB used to extract the folding sequence ?

protocols.simple_moves.FragmentMover: [ ERROR ] It appears that the fragments go up to residue 15 while the pose only has 14 residues!

Error: [ ERROR ] Must call shutdown() when finished using job distributor!

Which looks like somehow the arginine is being lost, even though the phosphotyrosine appears to be handled correctly.

Does anyone know what is causing this behavior, and whether this is an appropriate approach to ab intio for phosphopeptides (barring differences in secondary structure from Y to pY)?

 

Thank you for your help!

Kent