Page 23 - U. Protein structure and function

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

a,b
b
Sang-Ok Lee, Chong-Kil Lee, Kyoung-Seok Ryu c,d* and Seung-Wook Chi a,e*
a Disease Target Structure Research Center, Division of Biomedical Research, KRIBB, Daejeon 34141, Republic of Korea, b College of Pharmacy, Chungbuk National University, Cheongju 28644, Republic of Korea, c Protein
Structure Research Group, Korea Basic Science Institute, 162 Yeongudanji-ro, Ochang-eup, Cheongju-si, Chungcheongbuk-do 28119,Republic of Korea, d Department of Bio-Analytical Science, KBSI School of Bio-
Analytical Science, University of Science and Technology, 217 Gajeong-ro, Yuseong-gu, Daejeon 34113, Republic of Korea, and e Department of Proteome Structural Biology, KRIBB School of Bioscience, University of
Science and Technology, 217 Gajeong-ro, Yuseong-gu,Daejeon 34113, Republic of Korea.
BACKGROUND AIM
<Mitochondrial E3 ubiquitin ligase 1 (MUL1)>
First, we aim to identify functional
•The ubiquitylation activity of MUL1 is E2 enzyme for the ubiquitylation
important for the regulation of mitochondrial of the p53 transactivation domain
dynamics (fusion and fission), including (p53-TAD) by MUL1-RING
mitophagy, and is also involved in various domain.
biological processes such as apoptosis and
cell growth. Second, we are to perform the
•The presence of MUL1-RING is critical for purification and crystallization of
both the ubiquitylation and SUMOylation the MUL1-RING domain and the
activities of MUL1. MUL1-RING-Ube2D2 complex for
structure determination.
[Peng et al., Mitochondrion, 2016 ]
RESULTS

<An in vitro ubiquitylation assay by MUL1-RING alone> <X-ray diffraction and data-collection statistics for MUL1-
RING and the MUL1-Ube2D2 complex >

<SEC elution profile of the MUL1-RING-Ube2D2 mixture>

<MUL1-RING domain interacts mainly with <X-ray diffraction patterns of crystals of MUL1-RING and of
MUL1-RING complexed with Ube2D2>
p53-TAD2 subdomain>

CONCLUSION REFERENCES

1. We successfully purified and crystallized MUL1- Sang-Ok Lee et al., Acta Cryst F Structural Biology Communications,
RING and MUL1-RING-Ube2D2 complex. (2020). F76, 1-7.
2. We found that Ube2D2 is a functional E2
enzyme for the ubiquitylation of the p53 ACKNOWLEDGEMENTS
transactivation domain (p53-TAD) by MUL1- This work was supported by NRF grants funded by the Korean government (MSIP)
RING. (NRF-2017R1E1A1A01074403, NRF-2019M3E5D4069903 and NRF-
3. The 3D structures of MUL1-RING and the 2019M3A9C4076156) and by the KRIBB Research Initiative Program.
MUL1-RING–Ube2D2 complex are likely to
provide a structural basis for understanding the Contact information
unique ubiquitylation mechanism of MUL1-RING
alone that distinguishes it from other RING-E3 Seung-Wook Chi, Ph.D.
proteins. TEL : +82-42-860-4277, E-mail : swchi@kribb.re.kr