The RING domain of mitochondrial E3 ubiquitin ligase 1 and its complex with Ube2D2: crystallization and X-ray diffraction.

Publisher: John Wiley & Sons Inc Country of Publication: United States NLM ID: 101620319 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2053-230X (Electronic) Linking ISSN: 2053230X NLM ISO Abbreviation: Acta Crystallogr F Struct Biol Commun Subsets: MEDLINE

Original Publication: Malden, MA : John Wiley & Sons Inc., [2014]-

RING Finger Domains*; Mitochondria/*enzymology ; Ubiquitin-Conjugating Enzymes/*chemistry ; Ubiquitin-Protein Ligases/*chemistry; Crystallization ; Crystallography, X-Ray ; Gene Expression/genetics ; Humans ; Models, Molecular ; Protein Binding ; Tumor Suppressor Protein p53/chemistry ; Ubiquitin/chemistry ; Ubiquitination ; X-Ray Diffraction

Mitochondrial E3 ubiquitin ligase 1 (MUL1) is located in the mitochondrial outer membrane and regulates various biological processes, including apoptosis, cell growth, mitophagy and mitochondrial dynamics. The C-terminal region of MUL1 faces the cytoplasm and contains the RING domain (MUL1-RING) where the Ub~E2 thioester binds. Unlike most RING-type E3 enzymes, MUL1-RING alone does not have an additional region that recruits a substrate protein, yet is still able to ubiquitylate the substrate, the p53 protein. Nevertheless, the exact mechanism of the ubiquitylation of p53 by MUL1-RING has not yet been elucidated. In order to understand this novel ubiquitylation mechanism, it is necessary to determine the three-dimensional structures of MUL1-RING and of its complex with the cognate E2 enzyme. Here, Ube2D2 was validated as a functional E2 enzyme for the ubiquitylation of the p53 transactivation domain (p53-TAD) by MUL1-RING, and purification and crystallization processes for MUL1-RING and the MUL1-RING-Ube2D2 complex are reported.

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NRF-2017R1E1A1A01074403 National Research Foundation of Korea; NRF-2019M3E5D4069903 National Research Foundation of Korea; NRF-2019M3A9C4076156 National Research Foundation of Korea; T39632 Korea Basic Science Institute

Keywords: MUL1; MUL1-RING; RING domain; Ube2D2; crystallization; mitochondrial E3 ubiquitin ligase 1; ubiquitylation

0 (Tumor Suppressor Protein p53)
0 (Ubiquitin)
EC 2.3.2.23 (UBE2D2 protein, human)
EC 2.3.2.23 (Ubiquitin-Conjugating Enzymes)
EC 2.3.2.27 (MUL1 protein, human)
EC 2.3.2.27 (Ubiquitin-Protein Ligases)

Date Created: 20200114 Date Completed: 20200609 Latest Revision: 20240327

20250114

PMC6957114

10.1107/S2053230X19015395

31929179