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ZNFX1 activity-based probe crosslinking mass spectrometry

ABSTRACT: The aim of this experiment was to identify the catalytic cysteine in the antiviral E3 ligase ZNFX1 using activity-based probe (ABP)–enabled cross-linking mass spectrometry (XL-MS). A biotinylated E2~ubiquitin activity-based probe containing a cysteine-electrophile trap was used to covalently capture ubiquitin a transfer intermediate between UBE2D2~Ub and full-length human ZNFX1. Cross-linked peptide mapping was performed to identify probe-labelled residues and putative catalytic site.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Satpal Virdee

LAB HEAD: Satpal Virdee

PROVIDER: PXD073419 | Pride | 2026-05-22

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	FL_20220418_Dan_SV_20220314_ZNFX1_XL_5.raw	Raw
	ZNFX1_XLMS_Results_insol_5.xlsx	Xlsx
	checksum.txt	Txt

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Publications

ZNFX1 uses two-component ubiquitin circuitry to quarantine viral RNA.

Squair Daniel R DR Rivers Eilidh E Sowar Hanna H Balci Arda A Harmo Roosa R Wright David J DJ Beniwal Gaurav G Soetens Mathieu M Mathur Sunil S Tollervey Aidan A Wood Nicola T NT Pao Kuan-Chuan KC Stanton Callum C Fletcher Adam J AJ Virdee Satpal S

Molecular cell 20260301 6

The detection of viral RNA inside cells triggers a diverse range of antiviral responses, including global translation inhibition, interferon secretion, and RNA sequestration. Mutations in the gene zinc-finger NFX1-type containing 1 (ZNFX1) cause severe pediatric immunodeficiencies, including chronic viral infection and autoinflammation. Here, we show that ZNFX1 is an RNA helicase with cryptic and unusual bifurcating E3 ubiquitin ligase activity. Nucleotide-dependent RNA binding stimulates ZNFX1 ...[more]

PMID: 41861786

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