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Screening for covalent SUMOylation and noncovalent SUMO interactions with SIM-binding groove SUMO mutants

ABSTRACT: Small ubiquitin-like modifier (SUMO) proteins affect protein function through both covalent modification of target proteins and noncovalent protein-protein interactions. The best characterized mode of SUMO interaction involves binding of a SUMO-interaction motif (SIM) to a conserved binding groove in SUMO. Our knowledge on other types of SUMO interactions is still limited. Using SIM-binding groove SUMO2/3 mutants, we performed a pulldown approach to capture proteins capable of binding these SUMOs and thereby specifically enrich for proteins that bind to SUMO in an alternate manner, followed by mass spectrometry. Many proteins were able to bind both SUMO2/3 wildtype and the mutants, suggesting that the prevalence and relevance of SUMO-SIM-independent modes of SUMO interaction is currently underestimated. Using domain enrichment analysis, a group of WD40 repeat domain containing proteins were identified as novel class of SIM-independent SUMO interactors and we validated direct binding of SEC13 and SEH1L to SUMO in vitro. Additionally, we extended our understanding of the SUMO interactome to also include covalent SUMO modification in the context of the SIM-binding groove, uncovering its importance for covalent SUMO modification. Altogether, our dataset is a unique resource and offers valuable new insights on the intricacies of the SUMO interactome and SUMO targets.

INSTRUMENT(S): Orbitrap Exploris 480

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Laura Claessens

LAB HEAD: Alfred C.O.Vertegaal

PROVIDER: PXD050651 | Pride | 2025-04-03

REPOSITORIES: Pride

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			Action	DRS
	Covalentscreen_mqpar.xml	Xml
	Covalentscreen_samplenumbers.xlsx	Xlsx
	Covalentscreen_txt.zip	Other
	E20222002123a.raw	Raw
	E20222002124a.raw	Raw

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