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import os
import numpy as np
import sys
from timeit import default_timer as timer
import argparse
import glob
import uuid
import jax
import jax.numpy as jnp
from jax.lib import xla_bridge
from alphafold.common import residue_constants
from alphafold.common import protein
from alphafold.common import confidence
from alphafold.data import pipeline
from alphafold.model import data
from alphafold.model import config
from alphafold.model import model
import af2_util
parent = os.path.dirname(os.path.dirname(os.path.abspath(__file__)))
sys.path.append(os.path.join(parent, 'include'))
from silent_tools import silent_tools
from pyrosetta import *
from rosetta import *
init( '-in:file:silent_struct_type binary -mute all' )
def range1(size): return range(1, size+1)
#################################
# Parse Arguments
#################################
parser = argparse.ArgumentParser()
# I/O Arguments
parser.add_argument( "-pdbdir", type=str, default="", help='The name of a directory of pdbs to run through the model' )
parser.add_argument( "-silent", type=str, default="", help='The name of a silent file to run through the model' )
parser.add_argument( "-outpdbdir", type=str, default="outputs", help='The directory to which the output PDB files will be written. Only used when -pdbdir is active' )
parser.add_argument( "-outsilent", type=str, default="out.silent", help='The name of the silent file to which output structs will be written. Only used when -silent is active' )
parser.add_argument( "-runlist", type=str, default='', help="The path of a list of pdb tags to run. Only used when -pdbdir is active (default: ''; Run all PDBs)" )
parser.add_argument( "-checkpoint_name", type=str, default='check.point', help="The name of a file where tags which have finished will be written (default: check.point)" )
parser.add_argument( "-scorefilename", type=str, default='out.sc', help="The name of a file where scores will be written (default: out.sc)" )
parser.add_argument( "-maintain_res_numbering", action="store_true", default=False, help='When active, the model will not renumber the residues when bad inputs are encountered (default: False)' )
parser.add_argument( "-debug", action="store_true", default=False, help='When active, errors will cause the script to crash and the error message to be printed out (default: False)')
# AF2-Specific Arguments
parser.add_argument( "-max_amide_dist", type=float, default=3.0, help='The maximum distance between an amide bond\'s carbon and nitrogen (default: 3.0)' )
parser.add_argument( "-recycle", type=int, default=3, help='The number of AF2 recycles to perform (default: 3)' )
parser.add_argument( "-no_initial_guess", action="store_true", default=False, help='When active, the model will not use an initial guess (default: False)' )
parser.add_argument( "-force_monomer", action="store_true", default=False, help='When active, the model will predict the structure of a monomer (default: False)' )
args = parser.parse_args()
class FeatureHolder():
'''
This is a struct which holds the features for a single structure being run through the model
'''
def __init__(self, pose, monomer, binderlen, tag):
self.pose = pose
self.tag = tag
self.outtag = self.tag + '_af2pred'
self.seq = pose.sequence()
self.binderlen = binderlen
self.monomer = monomer
# Pre model features
self.initial_all_atom_positions = None
self.initial_all_atom_masks = None
# Post model features
self.outpose = None
self.plddt_array = None
self.score_dict = None
class AF2_runner():
'''
This class handles generating features, running the model, and parsing outputs
'''
def __init__(self, args, struct_manager):
self.max_amide_dist = args.max_amide_dist
# For timing
self.t0 = None
self.struct_manager = struct_manager
# Other models may be run but their weights will also need to be downloaded
self.model_name = "model_1_ptm"
model_config = config.model_config(self.model_name)
model_config.data.eval.num_ensemble = 1
model_config.data.common.num_recycle = args.recycle
model_config.model.num_recycle = args.recycle
model_config.model.embeddings_and_evoformer.initial_guess = False if args.no_initial_guess else True
model_config.data.common.max_extra_msa = 5
model_config.data.eval.max_msa_clusters = 5
package_root = os.path.dirname(os.path.dirname(os.path.dirname(os.path.realpath(__file__))))
params_dir = os.path.join(package_root, 'weight', 'AlphaFold2')
model_params = data.get_model_haiku_params(model_name=self.model_name, data_dir=params_dir)
self.model_runner = model.RunModel(model_config, model_params)
def featurize(self, feat_holder) -> None:
all_atom_positions, all_atom_masks = af2_util.af2_get_atom_positions(feat_holder.pose, self.struct_manager.tmp_fn)
feat_holder.initial_all_atom_positions = all_atom_positions
feat_holder.initial_all_atom_masks = all_atom_masks
initial_guess = af2_util.parse_initial_guess(feat_holder.initial_all_atom_positions)
# Determine which residues to template
if feat_holder.monomer:
# For monomers predict all residues
feat_holder.residue_mask = [False for i in range(len(feat_holder.seq))]
else:
# For interfaces fix the target and predict the binder
feat_holder.residue_mask = [int(i) > feat_holder.binderlen for i in range(len(feat_holder.seq))]
template_dict = af2_util.generate_template_features(
feat_holder.seq,
feat_holder.initial_all_atom_positions,
feat_holder.initial_all_atom_masks,
feat_holder.residue_mask
)
# Gather features
feature_dict = {
**pipeline.make_sequence_features(sequence=feat_holder.seq,
description="none",
num_res=len(feat_holder.seq)),
**pipeline.make_msa_features(msas=[[feat_holder.seq]],
deletion_matrices=[[[0]*len(feat_holder.seq)]]),
**template_dict
}
if feat_holder.monomer:
breaks = []
else:
breaks = af2_util.check_residue_distances(
feat_holder.initial_all_atom_positions,
feat_holder.initial_all_atom_masks,
self.max_amide_dist
)
feature_dict['residue_index'] = af2_util.insert_truncations(feature_dict['residue_index'], breaks)
feature_dict = self.model_runner.process_features(feature_dict, random_seed=0)
return feature_dict, initial_guess
def generate_scoredict(self, feat_holder, confidences, rmsds) -> None:
'''
Collect the confidence values, slicing them to the binder and target regions
then add the parsed scores to the score_dict
'''
binderlen = feat_holder.binderlen
plddt_array = confidences['plddt']
plddt = np.mean( plddt_array )
if feat_holder.monomer:
plddt_binder = np.mean( plddt_array )
plddt_target = float('nan')
else:
plddt_binder = np.mean( plddt_array[:binderlen] )
plddt_target = np.mean( plddt_array[binderlen:] )
pae = confidences['predicted_aligned_error']
if feat_holder.monomer:
pae_binder = np.mean( pae )
pae_target = float('nan')
pae_interaction_total = float('nan')
else:
pae_interaction1 = np.mean( pae[:binderlen,binderlen:] )
pae_interaction2 = np.mean( pae[binderlen:,:binderlen] )
pae_binder = np.mean( pae[:binderlen,:binderlen] )
pae_target = np.mean( pae[binderlen:,binderlen:] )
pae_interaction_total = ( pae_interaction1 + pae_interaction2 ) / 2
time = timer() - self.t0
score_dict = {
"plddt_total" : plddt,
"plddt_binder" : plddt_binder,
"plddt_target" : plddt_target,
"pae_binder" : pae_binder,
"pae_target" : pae_target,
"pae_interaction" : pae_interaction_total,
"binder_aligned_rmsd": rmsds['binder_aligned_rmsd'],
"target_aligned_rmsd": rmsds['target_aligned_rmsd'],
"time" : time
}
# Store this in the feature holder for later use
feat_holder.score_dict = score_dict
# If we ever want to write strings to the score file we can do it here
string_dict = None
self.struct_manager.record_scores(feat_holder.outtag, score_dict, string_dict)
print(score_dict)
print(f"Tag: {feat_holder.outtag} reported success in {time} seconds")
def process_output(self, feat_holder, feature_dict, prediction_result) -> None:
'''
Take the AF2 output, parse the confidence scores from this and register the scores in the score file
Also write out the structure
'''
# First extract the structure and confidence scores from the prediction result
structure_module = prediction_result['structure_module']
this_protein = protein.Protein(
aatype=feature_dict['aatype'][0],
atom_positions=structure_module['final_atom_positions'][...],
atom_mask=structure_module['final_atom_mask'][...],
residue_index=feature_dict['residue_index'][0] + 1,
b_factors=np.zeros_like(structure_module['final_atom_mask'][...]) )
confidences = {}
confidences['distogram'] = prediction_result['distogram']
confidences['plddt'] = confidence.compute_plddt(
prediction_result['predicted_lddt']['logits'][...])
if 'predicted_aligned_error' in prediction_result:
confidences.update(confidence.compute_predicted_aligned_error(
prediction_result['predicted_aligned_error']['logits'][...],
prediction_result['predicted_aligned_error']['breaks'][...]))
feat_holder.plddt_array = confidences['plddt']
# Calculate the RMSDs
target_mask = np.zeros(len(feat_holder.seq), dtype=bool)
target_mask[feat_holder.binderlen:] = True
rmsds = af2_util.calculate_rmsds(
feat_holder.initial_all_atom_positions,
this_protein.atom_positions,
target_mask
)
# Write the structure as a pdb file so Rosetta can read it
unrelaxed_pdb_lines = protein.to_pdb(this_protein)
with open(self.struct_manager.tmp_fn, 'w') as f: f.write(unrelaxed_pdb_lines)
# Now read the structure to a Rosetta pose
feat_holder.outpose = pyrosetta.pose_from_file(self.struct_manager.tmp_fn)
os.remove(self.struct_manager.tmp_fn)
# Now we can finally write the scores and the predicted structure to disk
self.generate_scoredict(feat_holder, confidences, rmsds)
self.struct_manager.dump_pose(feat_holder)
def process_struct(self, tag) -> None:
# Start the timer
self.t0 = timer()
# Load the structure
pose, monomer, binderlen, usetag = self.struct_manager.load_pose(tag)
# Store the pose in the feature holder
feat_holder = FeatureHolder(pose, monomer, binderlen, usetag)
print(f'Processing struct with tag: {feat_holder.tag}')
# Generate features
feature_dict, initial_guess = self.featurize(feat_holder)
# Run model
start = timer()
print(f'Running {self.model_name}')
prediction_result = self.model_runner.apply( self.model_runner.params,
jax.random.PRNGKey(0),
feature_dict,
initial_guess )
print(f'Tag: {feat_holder.tag} finished AF2 prediction in {timer() - start} seconds')
# Process outputs
self.process_output(feat_holder, feature_dict, prediction_result)
class StructManager():
'''
This class handles all of the input and output for the AF2 model. It deals with silent files vs. pdbs,
checkpointing, and writing of outputs
Note: This class could be moved to a separate file and shared with ProteinMPNN
'''
def __init__(self, args):
self.args = args
self.maintain_res_numbering = args.maintain_res_numbering
self.score_fn = args.scorefilename
# Generate a random unique temporary filename
self.tmp_fn = f'tmp_{uuid.uuid4()}.pdb'
self.force_monomer = args.force_monomer
self.silent = False
if not args.silent == '':
self.silent = True
self.struct_iterator = silent_tools.get_silent_index(args.silent)['tags']
self.sfd_in = rosetta.core.io.silent.SilentFileData(rosetta.core.io.silent.SilentFileOptions())
self.sfd_in.read_file(args.silent)
self.sfd_out = core.io.silent.SilentFileData(args.outsilent, False, False, "binary", core.io.silent.SilentFileOptions())
self.outsilent = args.outsilent
self.pdb = False
if not args.pdbdir == '':
self.pdb = True
self.pdbdir = args.pdbdir
self.outpdbdir = args.outpdbdir
self.struct_iterator = glob.glob(os.path.join(args.pdbdir, '*.pdb'))
# Parse the runlist and determine which structures to process
if args.runlist != '':
with open(args.runlist, 'r') as f:
self.runlist = set([line.strip() for line in f])
# Filter the struct iterator to only include those in the runlist
self.struct_iterator = [struct for struct in self.struct_iterator if '.'.join(os.path.basename(struct).split('.')[:-1]) in self.runlist]
print(f'After filtering by runlist, {len(self.struct_iterator)} structures remain')
# Assert that either silent or pdb is true, but not both
assert(self.silent ^ self.pdb), f'Both silent and pdb are set to {args.silent} and {args.pdb} respectively. Only one of these may be active at a time'
# Setup checkpointing
self.chkfn = args.checkpoint_name
self.finished_structs = set()
if os.path.isfile(self.chkfn):
with open(self.chkfn, 'r') as f:
for line in f:
self.finished_structs.add(line.strip())
def input_check(self, pose, tag) -> bool:
'''
This function checks that the given pose is valid for AF2. It specifically
checks that all residue indices are unique
'''
seen_indices = set()
# Loop through the PDBinfo of the pose and check that all residue indices are unique
pdbinfo = pose.pdb_info()
for resi in range1(pose.size()):
residx = pdbinfo.number(resi)
if residx in seen_indices:
print( f"\nNon-unique residue indices detected for tag: {tag}. " +
"This will cause AF2 to yield garbage outputs." )
return False
seen_indices.add(residx)
return True
def record_checkpoint(self, tag):
'''
Record the fact that this tag has been processed.
Write this tag to the list of finished structs
'''
with open(self.chkfn, 'a') as f:
f.write(f'{tag}\n')
def iterate(self):
'''
Iterate over the silent file or pdb directory and run the model on each structure
'''
# Iterate over the structs and for each, check that the struct has not already been processed
for struct in self.struct_iterator:
if self.pdb:
tag = '.'.join(os.path.basename(struct).split('.')[:-1])
else:
tag = struct
if tag in self.finished_structs:
print(f'{tag} has already been processed. Skipping')
continue
yield struct
def record_scores(self, tag, score_dict, string_dict):
'''
Record the scores for this structure to the score file.
Args:
tag (str): The tag for this structure
score_dict (dict): A dictionary of numerical scores to record
string_dict (dict): A dictionary of string scores to record
'''
# Check whether the score file exists
write_header = False
if not os.path.isfile(self.score_fn):
write_header = True
af2_util.add2scorefile(tag, self.score_fn, write_header, score_dict, string_dict)
def dump_pose(self, feat_holder):
'''
Dump this pose to either a silent file or a pdb file depending on the input arguments
'''
if feat_holder.monomer:
pose = feat_holder.outpose
else:
# Insert chainbreaks into the pose
pose = af2_util.insert_Rosetta_chainbreaks(feat_holder.outpose, feat_holder.binderlen)
# Add the plddt scores as b-factors to the pose
info = pose.pdb_info()
for resi in range1(pose.size()):
info.add_reslabel(resi, f'{feat_holder.plddt_array[resi-1]}')
for atom_i in range1(pose.residue(resi).natoms()):
info.bfactor(resi, atom_i, feat_holder.plddt_array[resi-1])
# Assign the pose the updated pdb_info
pose.pdb_info(info)
if self.pdb:
# If the outpdbdir does not exist, create it
# If there are parents in the path that do not exist, create them as well
if not os.path.exists(self.outpdbdir):
os.makedirs(self.outpdbdir)
pdbfile = os.path.join(self.outpdbdir, feat_holder.outtag + '.pdb')
pose.dump_pdb(pdbfile)
if self.silent:
struct = self.sfd_out.create_SilentStructOP()
struct.fill_struct(pose, feat_holder.outtag)
# Write the scores to the silent file
for scorename, value in feat_holder.score_dict.items():
struct.add_energy(scorename, value, 1)
self.sfd_out.add_structure(struct)
self.sfd_out.write_silent_struct(struct, self.outsilent)
def load_pose(self, tag):
'''
Load a pose from either a silent file or a pdb file depending on the input arguments.
Also run input checking on the pose to ensure that it is valid for AF2. The pose
will be automatically renumbered if -maintain_res_numbering is not False
'''
if not self.pdb and not self.silent:
raise Exception('Neither pdb nor silent is set to True. Cannot load pose')
if self.pdb:
pose = pose_from_pdb(tag)
usetag = '.'.join(os.path.basename(tag).split('.')[:-1])
if self.silent:
pose = Pose()
usetag = tag
self.sfd_in.get_structure(tag).fill_pose(pose)
# Run input checking on the pose
passes_check = self.input_check(pose, tag)
if not passes_check:
if not self.maintain_res_numbering:
print( f"Renumbering {tag}" )
info = core.pose.PDBInfo(pose)
pose.pdb_info(info)
else:
raise Exception( f"Pose {tag} failed input checking.")
# Determine whether we are working with a monomer or a complex
splits = pose.split_by_chain()
if len(splits) > 2:
raise Exception( f"Pose {tag} has more than two chains. This is not supported by this script." )
elif len(splits) < 1:
raise Exception( f"Pose {tag} is empty. This is not supported by this script." )
elif len(splits) == 1:
monomer = True
binderlen = -1
elif len(splits) == 2:
if self.force_monomer:
print( "/" * 60 )
print( f"Pose {tag} has two chains. But force_monomer is set to True. Treating as monomer.")
print( f"I am going to assume that the first chain is the binder and that is the chain I will predict")
print( "/" * 60 )
monomer = True
pose = splits[1]
binderlen = -1
else:
monomer = False
binderlen = splits[1].size()
return pose, monomer, binderlen, usetag
####################
####### Main #######
####################
device = xla_bridge.get_backend().platform
if device == 'gpu':
print('/' * 60)
print('/' * 60)
print('Found GPU and will use it to run AF2')
print('/' * 60)
print('/' * 60)
print('\n')
else:
print('/' * 60)
print('/' * 60)
print('WARNING! No GPU detected running AF2 on CPU')
print('/' * 60)
print('/' * 60)
print('\n')
struct_manager = StructManager(args)
af2_runner = AF2_runner(args, struct_manager)
for pdb in struct_manager.iterate():
if args.debug: af2_runner.process_struct(pdb)
else: # When not in debug mode the script will continue to run even when some poses fail
t0 = timer()
try: af2_runner.process_struct(pdb)
except KeyboardInterrupt: sys.exit( "Script killed by Control+C, exiting" )
except:
seconds = int(timer() - t0)
print( "Struct with tag %s failed in %i seconds with error: %s"%( pdb, seconds, sys.exc_info()[0] ) )
# We are done with one pdb, record that we finished
struct_manager.record_checkpoint(pdb)
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