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Ubiquitination sites on MIDN identified by mass spectrometry analysis

ABSTRACT: In E. coli, the pACYC-HA-Ub-E1 (UBA1)-UbcH5A-RNF126 and pET-22b-MIDN-His₆ plasmids were co-transformed. After IPTG induction to express all proteins required for ubiquitination, His-tagged MIDN was enriched with Ni-NTA resin and subjected to mass-spectrometric mapping of its ubiquitin-modified sites.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: yun yang

LAB HEAD: yun yang

PROVIDER: PXD069387 | Pride | 2026-06-01

REPOSITORIES: Pride

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			Action	DRS
	YS_CHI_532_126_MIDN.raw	Raw
	checksum.txt	Txt
	proteins.csv	Csv

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Publications

RNF126 writes a non-canonical ubiquitin code on midnolin to tune protein stability.

Yang Yun Y Ren Jin J Qiu Xiang X Liu Yanlin Y Yuan Shilin S Hu Ronggui R Xia Zhixiong Z Li Chuanyin C

Acta biochimica et biophysica Sinica 20260107 5

Midnolin (MIDN) is a newly recognized master regulator that drives ubiquitin-independent proteasomal degradation, yet the mechanisms governing its own turnover remain enigmatic. Here, we demonstrate that MIDN is ubiquitinated and identify RNF126 as the cognate E3 ligase. RNF126 physically associates with MIDN and catalyzes its ubiquitination, and mass spectrometry mapping reveals that this process occurs primarily at non-canonical cysteine, serine, and threonine residues (C230, C236, S237, T239, ...[more]

PMID: 41496599

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