Configuring Sage

Usage: sage [OPTIONS] <parameters> [mzml_paths]...
 
🔮 Sage 🧙 - Proteomics searching so fast it feels like magic!
 
Arguments:
  <parameters>     Path to configuration parameters (JSON file)
  [mzml_paths]...  Paths to mzML files to process. Overrides mzML files listed in the configuration file.
 
Options:
  -f, --fasta <fasta>
          Path to FASTA database. Overrides the FASTA file specified in the configuration file.
  -o, --output_directory <output_directory>
          Path where search and quant results will be written. Overrides the directory specified in the configuration file.
      --batch-size <batch-size>
          Number of files to search in parallel (default = number of CPUs/2)
      --parquet
          Write parquet files instead of tab-separated files
      --annotate-matches
          Write matched fragments output file.
      --write-pin
          Write percolator-compatible `.pin` output files
      --disable-telemetry-i-dont-want-to-improve-sage
          Disable sending telemetry data
  -h, --help
          Print help information
  -V, --version
          Print version information

# Sage will print additional information to the terminal
export SAGE_LOG=trace
# Sage will only use N threads, default is to use the number of logical CPU cores
export RAYON_NUM_THREADS=2
 
sage config.json

SAGE_LOG=trace RAYON_NUM_THREADS=2 sage config.json

# Reproduce the same results!
sage results.json

{
  "database": {
    // How many fragments are in each internal mass bucket
    // Use a lower value (8192) for high-res MS/MS, and higher values for low-res MS/MS
    "bucket_size": 32768,
    // This section is optional. Default is trypsin, using the parameters below
    "enzyme": {
      // Optional[int] {default=1}, Number of missed cleavages to allow
      "missed_cleavages": 2,
      // Optional[int] {default=5}, Minimum AA length of peptides to search
      "min_len": 5,
      // Optional[int] {default=50}, Maximum AA length of peptides to search
      "max_len": 50,
      // Optional[str] {default='KR'}. Amino acids to cleave at
      "cleave_at": "KR",
      // Optional[char/single AA] {default='P'}. Do not cleave if this AA follows the cleavage site
      "restrict": "P",
      // Optional[bool] {default=true}. Cleave at c terminus of matching amino acid
      "c_terminal": true
    },
    // Optional[float] {default=150.0}, Minimum mass of fragments to search
    "fragment_min_mz": 200.0,
    // Optional[float] {default=2000.0}, Maximum mass of fragments to search 
    "fragment_max_mz": 2000.0,
    // Optional[float] {default=500.0}, Minimum monoisotopic mass of peptides to fragment
    "peptide_min_mass": 500.0,
    // Optional[float] {default=5000.0}, Maximum monoisotopic mass of peptides to fragment
    "peptide_max_mass": 5000.0,
    // Optional[List[str]] {default=["b","y"]} Which fragment ions to generate and search?
    "ion_kinds": ["b", "y"],
    // Optional[int] {default=2}, Do not generate b1/b2/y1/y2 ions for preliminary searching.
    // Does not affect full scoring of PSMs!
    "min_ion_index": 2,
    // Optional[Dict[char, float]] {default=null}, static modifications
    "static_mods": {
      // Apply static modification to N-terminus of peptide
      "^": 304.207,
      // Apply static modification to lysine
      "K": 304.207,
      // Apply static modification to cysteine
      "C": 57.0215
    },
    // Optional[Dict[char, list[float]]] {default=null}, variable modifications
    "variable_mods": {
      // Variable mods are applied *before* static mod
      "M": [15.9949],
      "^Q": [-17.026549],
      // Applied to N-terminal glutamic acid
      "^E": [-18.010565],
      // Applied to peptide C-terminus
      "$": [49.2, 22.9],
      // Applied to protein N-terminus
      "[": 42.0,
      // Applied to protein C-terminus
      "]": 111.0
    },
    // Optional[int] {default=2} Limit k-combinations of variable modifications
    "max_variable_mods": 2,
    // Optional[str] {default="rev_"}: See notes above
    "decoy_tag": "rev_",
    // Optional[bool] {default="true"}: Ignore decoys in FASTA database matching `decoy_tag`
    "generate_decoys": false,
    // str: mandatory path to FASTA file
    "fasta": "dual.fasta"
  },
  // Optional - specify only if TMT or LFQ
  "quant": {
    // Optional[str] {default=null}, one of "Tmt6", "Tmt10", "Tmt11", "Tmt16", or "Tmt18"
    "tmt": "Tmt16",
    "tmt_settings": {
      // Optional[int] {default=3}, MS-level to perform TMT quantification on
      "level": 3,
      // Optional[bool] {default=false}, use Signal/Noise instead of intensity for TMT quant
      // Requires noise values in mzML
      "sn": false
    },
    // Optional[bool] {default=null}, perform label-free quantification
    "lfq": true,
    "lfq_settings": {
      // See documentation for details - recommend that you do not change this setting
      "peak_scoring": "Hybrid",
      // Optional["Sum" | "Apex"] {default="Sum"}, use sum or peak of MS1 traces in peak
      "integration": "Sum",
      // Optional[float] {default = 0.7}, normalized spectral angle (vs. theoretical isotopic envelope)
      // cutoff for calling an MS1 peak
      "spectral_angle": 0.7,
      // Optional[float] {default = 5.0}, tolerance for DICE window around calculated precursor mass
      "ppm_tolerance": 5.0
    }
  },
  // Tolerance can be either "ppm" or "da"
  "precursor_tol": {
    "da": [
      // This value is substracted from the experimental precursor to match theoretical peptides
      -500,
      // This value is added to the experimental precursor to match theoretical peptides
      100
    ]
  },
  "fragment_tol": {
    "ppm": [
     // This value is subtracted from the experimental fragment to match theoretical fragments 
     -10,
     // This value is added to the experimental fragment to match theoretical fragments 
     10
    ]
  },
  // Optional[Tuple[int, int]] {default=[2, 4]}
  // If charge states are not annotated in the mzML, or if `wide_window` mode is turned on, then consider
  // all precursors at z=2, z=3, z=4
  "precursor_charge": [2, 4],
  // Optional[Tuple[int, int]] {default=[0,0]}: C13 isotopic envelope to consider for precursor
  "isotope_errors": [
    // Consider -1 C13 isotope
    -1,
    // Consider up to +3 C13 isotope (-1/0/1/2/3) 
    3
  ],
  // Optional[bool] {default=false}: perform deisotoping and charge state deconvolution on MS2 spectra
  "deisotope": false,
  // Optional[bool] {default=false}: search for chimeric/co-fragmenting PSMs
  "chimera": false,
  // Optional[bool] {default=false}: _ignore_ `precursor_tol` and search in wide-window/DIA mode
  "wide_window": false,
  // Optional[bool] {default=true}: use retention time prediction model as an feature for LDA
  "predict_rt": false,
  // Optional[int] {default=15}: only process MS2 spectra with at least N peaks
  "min_peaks": 15,
  // Optional[int] {default=150}: take the top N most intense MS2 peaks to search,
  "max_peaks": 150,
  // Optional[int] {default=4}: minimum # of matched b+y ions to use for reporting PSMs
  "min_matched_peaks": 6,
  // Optional[int] {default=null}: maximum fragment ion charge states to consider,
  "max_fragment_charge": 1,
  // Optional[int] {default=1}: number of PSMs to report for each spectra. Higher values might disrupt PSM rescoring.
  "report_psms": 1,
  // Optional[str] {default=`.`}: Place output files in a given directory or S3 bucket/prefix
  "output_directory": "s3://bucket/prefix",
  // List[str]: representing paths to mzML (or gzipped-mzML) files for search
  "mzml_paths": [
    "local/path.mzML",
    "s3://bucket/PXD0000001/foo.mzML.gz"
  ]       
}