Protocol for SUMOylome analysis using a SUMO-T86K mutant combined with Lys-C digestion.

Yaning Wu, Rin Imai, Hidetaka Kosako, Haruhiko Siomi, Kensaku Murano

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Article in STAR protocols, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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5 · Who and what money

Authors and funding

5 authors.

Yaning WuDepartment of Molecular Biology, Keio University School of Medicine, Tokyo, Japan.

Rin ImaiDepartment of Molecular Biology, Keio University School of Medicine, Tokyo, Japan.

Hidetaka KosakoDivision of Cell Signaling, Institute of Advanced Medical Sciences, Tokushima University, Tokushima, Japan. Electronic address: kosako@tokushima-u.ac.jp.

Haruhiko SiomiNext Generation In Vivo Research Center (cNIVR), Chiba University, Chiba, Japan.

Kensaku MuranoDepartment of Molecular Biology, Keio University School of Medicine, Tokyo, Japan. Electronic address: kmurano@keio.jp.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Small ubiquitin-like modifier (SUMO) modification, also known as SUMOylation, is a critical post-translational modification. Here, we present a protocol using the SUMO-T86K (Thr86→Lys) mutant and Lys-C digestion, followed by enrichment of modified peptides and mass spectrometry, to identify SUMOylated proteins and modification sites. This strategy overcomes a major limitation of the commonly used SUMO-T86R (Thr86→Arg) approach with trypsin digestion, in which the resulting diglycine (GG) remnant is indistinguishable from ubiquitin modifications. For complete details on the use and execution of this protocol, please refer to Wu et al.

Indexed as

ProteomicsSmall Ubiquitin-Related Modifier ProteinsSumoylationHumansLysineMass SpectrometryMutationProtein Processing, Post-TranslationalProteomeLysineProteomeSmall Ubiquitin-Related Modifier ProteinsCell-based AssaysGene ExpressionMass SpectrometryMolecular BiologyProtein BiochemistryProteomics

Identifiers

PMID42001423

DOI10.1016/j.xpro.2026.104510

PMCPMC13103552

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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.