Affiliation:
1. School of Pharmacy, Hangzhou Normal University, Hangzhou, Zhejiang, China
2. Key Laboratory of Elemene Class
Anti-Cancer Chinese Medicines, Engineering Laboratory of Development and Application of Traditional Chinese
Medicines, Collaborative Innovation Center of Traditional Chinese Medicines of Zhejiang Province, Hangzhou Normal University, Hangzhou, Zhejiang, China
Abstract
Background:
UDP-glucuronosyltransferases (UGTs) play a crucial role in maintaining
endobiotic homeostasis and metabolizing xenobiotic compounds, particularly clinical drugs. However,
the detailed catalytic mechanism of UGTs has not been fully elucidated due to the limited
availability of reliable protein structures. Determining the catalytic domain of human UGTs has
proven to be a significant challenge, primarily due to the difficulty in purifying and crystallizing
the full-length protein.
Objective:
This study focused on the human UGT2B10 C-terminal cofactor binding domain, aiming
to provide structural insights into the fundamental catalytic mechanisms.
Method:
In this study, the C-terminal sugar-donor binding domain of human UGT2B10 was purified
and crystallized using the vapor-diffusion method. The resulting UGT2B10 CTD crystals displayed
high-quality diffraction patterns, allowing for data collection at an impressive resolution of
1.53 Å using synchrotron radiation. Subsequently, the structure of the UGT2B10 CTD was determined
using the molecule replacement method with a homologous structure.
Results:
The crystals were monoclinic, belonging to the space C2 with unit-cell parameters a =
85.90 Å, b = 58.39 Å, c = 68.87 Å, α = γ = 90°, and β = 98.138°. The Matthews coefficient VM
was determined to be 2.24 Å3 Da-1 (solvent content 46.43%) with two molecules in the asymmetric
unit.
Conclusion:
The crystal structure of UGT2B10 CTD was solved at a high resolution of 1.53 Å, revealing
a conserved cofactor binding pocket. This is the first study determining the C-terminal cofactor
binding domain of human UGT2B10, which plays a key role in additive drug metabolism.
Funder
National Natural Science Foundation of China
National Science and Technology Innovation 2030 Major Project of the Ministry of Science and Technology of China
Publisher
Bentham Science Publishers Ltd.
Subject
Biochemistry,General Medicine,Structural Biology