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CLI Reference

mokume provides four main commands. Use mokume --help or mokume <command> --help for details.

features2proteins

The unified pipeline: features to protein quantification in one step.

mokume features2proteins [OPTIONS]

Required Options

Option Description
-p/--parquet Input parquet file (quantms.io/qpx format)
-o/--output Output protein intensities CSV

Input & Filtering

Option Default Description
-s/--sdrf none SDRF file for sample metadata
--min-aa 7 Minimum amino acid length
--min-unique 2 Minimum unique peptides per protein
--remove-contaminants on Remove contaminants and decoys
--keep-contaminants off Keep contaminants and decoys

Quantification

Option Default Description
--quant-method maxlfq Method: maxlfq, directlfq, ibaq, topn, sum, median, ratio
--fasta none FASTA file (required for iBAQ)
--topn 3 N for TopN quantification
--ion-alignment none Ion alignment: none or hierarchical
--directlfq-cores auto CPU cores for DirectLFQ
--directlfq-min-nonan 1 Min non-NaN values for DirectLFQ

Normalization

Option Default Description
--run-normalization median Run-level: median, mean, iqr, max, max_min, none
--sample-normalization globalMedian Sample-level: globalMedian, conditionMedian, hierarchical, none
--normalization-proteins none File with protein IDs for normalization

IRS (Multi-Plex TMT)

Option Default Description
--irs off Enable IRS normalization
--irs-reference-samples auto Comma-separated reference sample names
--irs-sdrf-column auto SDRF column for reference detection
--irs-sdrf-values auto Values indicating reference samples
--irs-reference-regex pool\|powder\|ref\|reference\|bridge Regex for reference auto-detection
--irs-stat median Plex reference statistic: median or mean
--irs-remove-reference off Remove reference samples from output

Coverage Filter

Option Default Description
--coverage-threshold none Min fraction of non-missing values per condition

Ratio Quantification

Option Default Description
--ratio-fraction-merge mean Fraction merge strategy: mean or max

Batch Correction

Option Default Description
--batch-correction off Enable ComBat batch correction
--batch-method sample_prefix Detection method: sample_prefix, run, column
--batch-covariates none Comma-separated SDRF columns to preserve

Differential Expression

Option Default Description
--de off Enable differential expression analysis
--de-contrasts all pairs Comma-separated contrasts (e.g., "A-B,A-C")
--de-method ttest Method: ttest or limma
--de-log2fc 0.5 Minimum absolute log2 fold change
--de-fdr 0.05 Maximum FDR threshold
--de-output auto Output file for DE results

Plots & Reports

Option Default Description
--plot-dir none Output directory for plots
--plot-volcano off Generate volcano plots
--plot-heatmap off Generate heatmaps
--plot-pca off Generate PCA plots
--highlight-genes none Comma-separated gene names to highlight
--interactive-report off Generate interactive HTML QC report
--report-output auto Output path for HTML report

Export

Option Default Description
--export-peptides none Export normalized peptides to file
--export-ions none Export normalized ions (DirectLFQ only)

features2peptides

Feature-level to peptide-level normalization.

mokume features2peptides [OPTIONS]

Core Options

Option Default Description
-p/--parquet required Input parquet file
-s/--sdrf none SDRF file for metadata
-o/--output required Output peptide intensity file
--min_aa 7 Minimum amino acid length
--min_unique 2 Minimum unique peptides per protein
--remove_ids none File with protein IDs to exclude
--remove_decoy_contaminants off Remove decoys and contaminants
--remove_low_frequency_peptides off Remove peptides in <20% of samples

Normalization

Option Default Description
--run-normalization median Feature normalization: mean, median, iqr, none
--sample-normalization globalMedian Sample normalization: globalMedian, conditionMedian, hierarchical, none
--skip_normalization off Skip all normalization
--log2 off Log2 transform output
--save_parquet off Save output as parquet

Filter Configuration

Option Default Description
--filter-config none YAML/JSON filter configuration file
--generate-filter-config none Generate example config and exit
--filter-min-intensity none Min intensity threshold (override)
--filter-cv-threshold none Max CV across replicates (override)
--filter-charge-states none Comma-separated charge states (override)
--filter-max-missed-cleavages none Max missed cleavages (override)
--filter-exclude-modifications none Comma-separated modifications (override)
--filter-min-unique-peptides none Min unique peptides (override)
--filter-min-features none Min features per run (override)
--filter-max-missing-rate none Max missing rate (override)

peptides2protein

Protein quantification from normalized peptide data.

mokume peptides2protein [OPTIONS]
Option Default Description
-p/--peptides required Input peptide intensity file
-f/--fasta none FASTA file (required for iBAQ)
--method ibaq Method: ibaq, top3, top5, top10, topn, maxlfq, sum, directlfq
-e/--enzyme Trypsin Enzyme for in-silico digestion
-n/--normalize off Normalize quantification values
--min_aa 7 Min amino acid length
--max_aa 30 Max amino acid length
-t/--tpa off Calculate TPA (iBAQ only)
-r/--ruler off ProteomicRuler (iBAQ only)
-i/--ploidy 2 Ploidy number
-m/--organism human Organism for histone data
-c/--cpc 200 Cellular protein concentration (g/L)
--topn_n 3 N for TopN quantification
--threads -1 Threads for MaxLFQ (-1 = all cores)
--min_nonan 1 Min non-NaN for DirectLFQ
-o/--output required Output file path
--verbose off Print distribution info
--qc_report QCprofile.pdf QC report PDF path

correct-batches

Standalone batch correction for pre-quantified data.

mokume correct-batches [OPTIONS]
Option Default Description
-f/--folder required Folder with TSV files
-p/--pattern *ibaq.tsv File matching pattern
-o/--output required Output file path
-sid/--sample_id_column SampleID Sample ID column
-pid/--protein_id_column ProteinName Protein ID column
-ibaq/--ibaq_raw_column IBAQ Raw intensity column
--ibaq_corrected_column IBAQ_BEC Corrected intensity column
--comment # Comment character
--sep \t Field separator
--export_anndata off Export to AnnData h5ad

tsne-visualization

t-SNE dimensionality reduction visualization.

mokume tsne-visualization [OPTIONS]
Option Default Description
-f/--folder required Folder with protein files
-o/--pattern proteins.tsv File matching pattern