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Identification of MARCH2 E3 ligase-substrates using proximity-dependent biotin labeling method

ABSTRACT: MARCH2 is known to be involved in intracellular vesicular trafficking. To identify its substrates, we have recently developed a method based on proximity-dependent biotin labelling. In this protocol, the MARCH2 ubiquitin ligase of interest is expressed as a fusion to Escherichia coli biotin ligase BirA together with a biotin acceptor peptide(AP)-tagged ubiquitin. The BirA-directed biotin labeling of AP depends on the proximity of the two fusion proteins in the cell, which leads to preferential labeling of ubiquitinated E3 substrates. In this study, we applied this procedure to MARCH2 and identified its substrates.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell, Uterine Cervix

DISEASE(S): Cervical Adenocarcinoma

SUBMITTER: Wonjin Yoo

LAB HEAD: Jong-Bok Yoon

PROVIDER: PXD013601 | Pride | 2019-06-06

REPOSITORIES: pride

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			Action	DRS
	MARCH2.mgf	Mgf
	MARCH2.raw	Raw

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Publications

The E3 ubiquitin ligase MARCH2 regulates ERGIC3-dependent trafficking of secretory proteins.

Yoo Wonjin W Cho Eun-Bee EB Kim Sungjoo S Yoon Jong-Bok JB

The Journal of biological chemistry 20190529 28

The E3 ubiquitin ligase membrane-associated ring-CH-type finger 2 (MARCH2) is known to be involved in intracellular vesicular trafficking, but its role in the early secretory pathway between the endoplasmic reticulum (ER) and Golgi compartments is largely unknown. Human ER-Golgi intermediate compartment protein 2 (ERGIC2) and ERGIC3 are orthologs of Erv41 and Erv46 in yeast, proteins that form a heteromeric complex, cycle between the ER and Golgi, and function as cargo receptors in both anterogr ...[more]

PMID: 31142615

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