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Here, we defined the Ac/N domain of the endoplasmic reticulum MARCHF6 E3 ubiquitin ligase through a systematic analysis of its cytosol-facing regions using alanine-stretch mutagenesis, chemical crosslinking-based co-immunoprecipitation–immunoblotting, and split-ubiquitin assays in human and yeast cells

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Protein crystallization and structure determination Rv3722c (~40 mg mL −1 ) was co-crystalized with ligands by sitting drop vapor diffusion at 17 °C