Proteomics

Dataset Information

55

Identifying the Ubiquitination Targets of E6AP by Orthogonal Ubiquitin Transfer


ABSTRACT: E3 ubiquitin (UB) ligases, the ending module of the E1-E2-E3 cascades, transmit diverse signals in the cell by attaching UB to cellular proteins. Identifying E3 substrates holds the key to elucidate the roles of E3s in cell regulation. We constructed an orthogonal UB transfer (OUT) cascade to identify the substrates of E6AP, a HECT E3 also known as Ube3a that is implicated in neurodevelopmental disorders and cancer. We used yeast cell surface display to engineer E6AP to exclusively transfer an affinity-tagged UB variant (xUB) to its substrate proteins. Proteomic identification of xUB-conjugated proteins in HEK293 cells afforded 131 potential E6AP targets. Among them we verified MAPK1, CDK1 and CDK4, and PRMT5 are directly ubiquitinated by E6AP in vitro and in the cell. Our work establishes OUT as an efficient platform to profile E6AP substrates and would guide the engineering of OUT cascades with other E3s to interrogate their biological functions.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo sapiens  

TISSUE(S): Cell Culture

DISEASE(S): Not Available

SUBMITTER: Duc Duong  

LAB HEAD: Jun Yin

PROVIDER: PXD005584 | Pride | 2018-02-07

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
HUMAN.fasta Fasta
ctl_01.msf Msf
ctl_01.pdResult Other
ctl_01.raw Raw
ctl_2and3.msf Msf
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Publications

Identifying the ubiquitination targets of E6AP by orthogonal ubiquitin transfer.

Wang Yiyang Y   Liu Xianpeng X   Zhou Li L   Duong Duc D   Bhuripanyo Karan K   Zhao Bo B   Zhou Han H   Liu Ruochuan R   Bi Yingtao Y   Kiyokawa Hiroaki H   Yin Jun J  

Nature communications 20171220 1


E3 ubiquitin (UB) ligases are the ending modules of the E1-E2-E3 cascades that transfer UB to cellular proteins and regulate their biological functions. Identifying the substrates of an E3 holds the key to elucidate its role in cell regulation. Here, we construct an orthogonal UB transfer (OUT) cascade to identify the substrates of E6AP, a HECT E3 also known as Ube3a that is implicated in cancer and neurodevelopmental disorders. We use yeast cell surface display to engineer E6AP to exclusively t  ...[more]

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