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Identification of a USP9X Substrate NFX1-123 by SILAC-based Quantitative Proteomics

ABSTRACT: The deubiquitinase USP9X related with multiple diseases including neurodegeneration, epilepsy and various type of tumor, by targeting different substrates. In our study, to deep study USP9X functional candidate substrates, we performed SILAC-based quantitative proteomic to compare knockdown USP9X with wild type HeLa cells to screen USP9X potential substrates. Further, we carried out Flag-NFX1-123 affinity tag-based interaction mass spectrometry method and verified that the X-box binding nuclear factor NFX1-123 is a substrate of USP9X. Further experimental evidences confirmed that USP9X stabilizedNFX1-123 protein level rather than mRNA levels via direct interaction and efficient deubiquitination of NFX1-123 protein

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Keratinocyte

DISEASE(S): Cervix Carcinoma

SUBMITTER: xiangling chen

LAB HEAD: Hu Zhou

PROVIDER: PXD012871 | Pride | 2019-05-13

REPOSITORIES: Pride

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Identification of a USP9X Substrate NFX1-123 by SILAC-Based Quantitative Proteomics.

Chen Xiangling X Lu Dayun D Gao Jing J Zhu Hongwen H Zhou Yanting Y Gao Daming D Zhou Hu H

Journal of proteome research 20190522 6

The deubiquitinase USP9X is involved in multiple diseases including neurodegeneration, epilepsy, and various types of tumors by targeting different substrates. In the present study, we aimed to explore the potential substrates of USP9X and performed SILAC-based quantitative proteomics to compare these substrates in USP9X-knockdown and wild-type HeLa cells. We consequently carried out Flag-NFX1-123 tag affinity-based mass spectrometry and confirmed that the X-box binding nuclear factor NFX1-123 i ...[more]

PMID: 31059266

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Loss of Usp9x increases Wnt and Notch signaling in neural progenitors

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