Evidence map›Paper›PMID 41559053›Full record

ArticleNature communications2026

Community benchmarking and evaluation of human unannotated microprotein detection by mass spectrometry based proteomics.

Aaron Wacholder, Eric W Deutsch, Leron W Kok, Jip T van Dinter, Jiwon Lee, James C Wright, Sebastien Leblanc, Ayodya H Jayatissa, Kevin Jiang, Ihor Arefiev and 24 more

Erratum issuedAbstract read
In one paragraph

Article in Nature communications, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 8 papers.

0numbers the graph read from it
0cells of the map it votes in
8citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

8 citing papers in PubMed.

  1. Review
  2. Article
  3. Article
  4. Article
  5. Article
  6. Review
  7. Article
  8. Article
4 · The record

Corrections and comments

5 · Who and what money

Authors and funding

34 authors.

Aaron WacholderDepartment of Computational and Systems Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA.ORCID http://orcid.org/0000-0001-8739-0029
Eric W DeutschInstitute for Systems Biology, Seattle, WA, USA.ORCID http://orcid.org/0000-0001-8732-0928
Leron W KokPrincess Máxima Center for Pediatric Oncology, Utrecht, The Netherlands.ORCID http://orcid.org/0009-0008-0841-2313
Jip T van DinterPrincess Máxima Center for Pediatric Oncology, Utrecht, The Netherlands.ORCID http://orcid.org/0000-0002-3749-1531
Jiwon LeeDepartment of Computational and Systems Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA.ORCID http://orcid.org/0000-0002-4079-7494
James C WrightFunctional Proteomics, Institute of Cancer Research, London, UK.ORCID http://orcid.org/0000-0001-6950-4328
Sebastien LeblancDepartment of Biochemistry and Functional Genomics, Université de Sherbrooke, Sherbrooke, QC, Canada.
Ayodya H JayatissaYale University Institute for Biomolecular Design and Discovery, West Haven, CT, USA.
Kevin JiangYale University Institute for Biomolecular Design and Discovery, West Haven, CT, USA.ORCID http://orcid.org/0000-0003-3258-4445
Ihor ArefievMedical Genetics Service, Pediatrics Department, University of Sherbrooke Cancer Research Institute, Sherbrooke, QC, Canada.
Kevin CaoDepartment of Pharmaceutical Sciences, University of California, Irvine, CA, USA.
Francis BourassaMedical Genetics Service, Pediatrics Department, University of Sherbrooke Cancer Research Institute, Sherbrooke, QC, Canada.ORCID http://orcid.org/0009-0001-9012-7245
Felix-Antoine TrifiroMedical Genetics Service, Pediatrics Department, University of Sherbrooke Cancer Research Institute, Sherbrooke, QC, Canada.
Michal Bassani-SternbergUniversity Hospital of Lausanne, Lausanne, Switzerland.ORCID http://orcid.org/0000-0002-1934-954X
Pavel V BaranovSchool of Biochemistry and Cell Biology, University College Cork, Cork, Ireland.
Annelies BogaertVIB Center for Medical Biotechnology, VIB, Ghent, Belgium.
Sonia ChothaniDuke-NUS Medical School, Singapore, Singapore.ORCID http://orcid.org/0000-0002-1010-7069
Ivo Fierro-MontiEuropean Molecular Biology Laboratory, European Bioinformatics Institute, Wellcome Genome Campus, Hinxton, Cambridge, UK.ORCID http://orcid.org/0000-0002-5460-2117
Daria FijalkowskaVIB Center for Medical Biotechnology, VIB, Ghent, Belgium.
Kris GevaertVIB Center for Medical Biotechnology, VIB, Ghent, Belgium.ORCID http://orcid.org/0000-0002-4237-0283
Norbert HubnerCardiovascular and Metabolic Sciences, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.ORCID http://orcid.org/0000-0002-1218-6223
Jonathan M MudgeEuropean Molecular Biology Laboratory, European Bioinformatics Institute, Wellcome Genome Campus, Hinxton, Cambridge, UK.ORCID http://orcid.org/0000-0003-4789-7495
Jorge Ruiz-OreraCardiovascular and Metabolic Sciences, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.ORCID http://orcid.org/0000-0002-8317-0034
Jana SchulzAltos Labs, Cambridge Institute of Science, Granta Park, Cambridge, UK.
Juan Antonio VizcaínoEuropean Molecular Biology Laboratory, European Bioinformatics Institute, Wellcome Genome Campus, Hinxton, Cambridge, UK.ORCID http://orcid.org/0000-0002-3905-4335
John R PrensnerDepartment of Pediatrics, Division of Pediatric Hematology/Oncology, University of Michigan Medical School, Ann Arbor, MI, USA.ORCID http://orcid.org/0000-0002-7024-636X
Marie A BrunetMedical Genetics Service, Pediatrics Department, University of Sherbrooke Cancer Research Institute, Sherbrooke, QC, Canada.ORCID http://orcid.org/0000-0001-5973-3522
Thomas F MartinezDepartment of Pharmaceutical Sciences, University of California, Irvine, CA, USA.ORCID http://orcid.org/0000-0002-4011-8164
Sarah A SlavoffYale University Institute for Biomolecular Design and Discovery, West Haven, CT, USA.ORCID http://orcid.org/0000-0002-4443-2070
Xavier RoucouDepartment of Biochemistry and Functional Genomics, Université de Sherbrooke, Sherbrooke, QC, Canada.ORCID http://orcid.org/0000-0001-9370-5584
Jyoti S ChoudharyFunctional Proteomics, Institute of Cancer Research, London, UK.ORCID http://orcid.org/0000-0003-0881-5477
Sebastiaan van HeeschPrincess Máxima Center for Pediatric Oncology, Utrecht, The Netherlands.
Robert L MoritzInstitute for Systems Biology, Seattle, WA, USA.ORCID http://orcid.org/0000-0002-3216-9447
Anne-Ruxandra CarvunisDepartment of Computational and Systems Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA. anc201@pitt.edu.ORCID http://orcid.org/0000-0002-6474-6413

Funding

GENCODE Resource ProjectU41HG007234 · NHGRI · SANGER INSTITUTE · PI FLICEK, PAUL · 2013 to 2020
$20.3M
Shortening the development cycle time of Trans Proteomic Pipeline tools with high performance computingR01GM087221 · NIGMS · INSTITUTE FOR SYSTEMS BIOLOGY · PI DEUTSCH, ERIC, MORITZ, ROBERT L · 2010 to 2021
$6.0M
Global proteomics mass spectrometry data sharing infrastructureR24GM148372 · NIGMS · UNIVERSITY OF CALIFORNIA, SAN DIEGO · PI Nuno Bandeira · 2023 to 2026
$2.8M
Predoctoral Training at the Interface Chemistry and BiologyT32GM149444 · NIGMS · YALE UNIVERSITY · PI Jason Michael Crawford · 2023 to 2026
$2.2M
Deciphering non-canonical translation in high risk medulloblastomaK08CA263552 · NCI · UNIVERSITY OF MICHIGAN AT ANN ARBOR · PI John Prensner · 2022 to 2026
$1.2M
Deciphering the Interactome and Functions of Disordered MicroproteinsR35GM157126 · NIGMS · UNIVERSITY OF CALIFORNIA-IRVINE · PI Thomas Farid Martinez · 2025 to 2026
$769k
Acquisition of Fusion Lumos Orbitrap mass spectrometerS10OD026936 · OD · INSTITUTE FOR SYSTEMS BIOLOGY · PI MORITZ, ROBERT L · 2019 to 2019
$600k
Human Frontier Science Program (HFSP) https://doi.org/10.52044/HFSP.RGP0042023.pc.gr.168590NCI NIH HHS K08 CA263552NHGRI NIH HHS U41 HG007234NIGMS NIH HHS R01 GM087221NIGMS NIH HHS R24 GM148372NIGMS NIH HHS R35 GM157126NIGMS NIH HHS T32 GM149444NIH HHS S10 OD026936Wellcome Trust
6 · The paper itself

Abstract

Thousands of short open reading frames (sORFs) are translated outside of annotated coding sequences. Recent studies have pioneered searching for sORF-encoded microproteins in mass spectrometry (MS)-based proteomics and peptidomics datasets. Here, we assessed literature-reported MS-based identifications of unannotated human proteins. We find that studies vary by three orders of magnitude in the number of unannotated proteins they report. Of nearly 10,000 reported sORF-encoded peptides, 96% were unique to a single study, and 12% mapped to annotated proteins or proteoforms. Manual curation of a benchmark dataset of 406 manually evaluated spectra from 204 sORF-encoded proteins revealed large variation in peptide-spectrum match (PSM) quality between studies, with immunopeptidomics studies generally reporting higher quality PSMs than conventional enzymatic digests of whole cell lysates. We estimate that 65% of predicted sORF-encoded protein detections in immunopeptidomics studies were supported by high-quality PSMs versus 7.8% in non-immunopeptidomics datasets. Our work stresses the need for standardized protocols and analysis workflows to guide future advancements in microprotein detection by MS towards uncovering how many human microproteins exist.

Indexed as

Mass SpectrometryMicropeptidesProteomicsBenchmarkingHumansMolecular Sequence AnnotationOpen Reading FramesMicropeptides

Identifiers

PMID41559053
PMCPMC12865046

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LicenceCC BY-NC-ND
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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.