Molecular basis for cysteine oxidation by plant cysteine oxidases from Arabidopsis thaliana

Zhenzhen Chen; Qiong Guo; Gao Wu; Jie Wen; Shanhui Liao; Chao Xu

doi:10.1016/j.jsb.2020.107663

Molecular basis for cysteine oxidation by plant cysteine oxidases from Arabidopsis thaliana

J Struct Biol. 2021 Mar;213(1):107663. doi: 10.1016/j.jsb.2020.107663. Epub 2020 Nov 15.

Authors

Zhenzhen Chen¹, Qiong Guo², Gao Wu³, Jie Wen⁴, Shanhui Liao⁵, Chao Xu⁶

Affiliations

¹ The First Affiliated Hospital of University of Science and Technology of China (Anhui Provincial Hospital), 230000 Hefei, China; MOE Key Laboratory for Membraneless Organelles and Cellular Dynamics, Hefei National Laboratory for Physical Sciences at the Microscale, University of Science and Technology of China, Hefei 230027, China.
² MOE Key Laboratory for Membraneless Organelles and Cellular Dynamics, Hefei National Laboratory for Physical Sciences at the Microscale, University of Science and Technology of China, Hefei 230027, China.
³ The First Affiliated Hospital of University of Science and Technology of China (Anhui Provincial Hospital), 230000 Hefei, China.
⁴ The First Affiliated Hospital of University of Science and Technology of China (Anhui Provincial Hospital), 230000 Hefei, China. Electronic address: jiewen@ustc.edu.cn.
⁵ MOE Key Laboratory for Membraneless Organelles and Cellular Dynamics, Hefei National Laboratory for Physical Sciences at the Microscale, University of Science and Technology of China, Hefei 230027, China. Electronic address: ajsod@mail.ustc.edu.cn.
⁶ MOE Key Laboratory for Membraneless Organelles and Cellular Dynamics, Hefei National Laboratory for Physical Sciences at the Microscale, University of Science and Technology of China, Hefei 230027, China. Electronic address: xuchaor@ustc.edu.cn.

PMID: 33207269
DOI: 10.1016/j.jsb.2020.107663

Abstract

Plant Cysteine Oxidases (PCOs) play important roles in controlling the stability of Group VII ethylene response factors (ERF-VIIs) via Arg/N-degron pathway through catalyzing the oxidation of their N-Cys for subsequent Arginyl-tRNA--protein transferase 1 (ATE1) mediated arginine installation. Here we presented the crystal structures of PCO2, PCO4, and PCO5 from Arabidopsis thaliana (AtPCOs) and examined their in vitro activity by Mass spectrometry (MS). On the basis of Tris-bound AtPCO2, we modelled the structure of Cys-bound AtPCO2 and identified key AtPCO2 residues involved in N-Cys recognition and oxidation. Alanine substitution of potential N-Cys interaction residues impaired the activity of AtPCO5 remarkably. The structural research, complemented by mutagenesis and MS experiments, not only uncovers the substrate recognition and catalytic mode by AtPCOs, but also sheds light on the future design of potent inhibitors for plant cysteine oxidases.

Keywords: Cysteine oxidation; Mass spectrometry; Plant Cysteine Oxidase(s); X-ray crystallography.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Arabidopsis / metabolism*
Arabidopsis Proteins / metabolism*
Arginine / metabolism
Cysteine / metabolism*
Cysteine Dioxygenase / metabolism*
Oxidation-Reduction

Substances

Arabidopsis Proteins
Arginine
Cysteine Dioxygenase
Cysteine