Effect of the metal ion-induced carbonylation modification of mitochondrial membrane channel protein VDAC on cell vitality, seedling growth and seed aging

Ying Li; Chang Liu; Manyao Qi; Tiantian Ye; Ying Kang; Yu Wang; Xiaofeng Wang; Hua Xue

doi:10.3389/fpls.2023.1138781

Effect of the metal ion-induced carbonylation modification of mitochondrial membrane channel protein VDAC on cell vitality, seedling growth and seed aging

Front Plant Sci. 2023 May 31:14:1138781. doi: 10.3389/fpls.2023.1138781. eCollection 2023.

Authors

Ying Li¹, Chang Liu¹, Manyao Qi¹, Tiantian Ye¹, Ying Kang¹, Yu Wang¹, Xiaofeng Wang¹, Hua Xue¹

Affiliation

¹ State Key Laboratory of Tree Genetics and Breeding, National Engineering Research Center of Tree Breeding and Ecological Remediation, College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing, China.

Abstract

Introduction: Seeds are the most important carrier of germplasm preservation. However, an irreversible decrease in vigor can occur after the maturation of seeds, denoted as seed aging. Mitochondrion is a crucial organelle in initiation programmed cell death during seed aging. However, the underlying mechanism remains unclear.

Methods: Our previous proteome study found that 13 mitochondria proteins underwent carbonylation modification during the aging of Ulmus pumila L. (Up) seeds. This study detected metal binding proteins through immobilized metal affinity chromatography (IMAC), indicating that metal binding proteins in mitochondria are the main targets of carbonization during seed aging. Biochemistry, molecular and cellular biology methods were adopted to detect metal-protein binding, protein modification and subcellular localization. Yeast and Arabidopsis were used to investigate the biological functions in vivo.

Results and discussion: In IMAC assay, 12 proteins were identified as Fe²+/Cu²+/Zn²+ binding proteins, including mitochondrial voltage dependent anion channels (VDAC). UpVDAC showed binding abilities to all the three metal ions. His204Ala (H204A) and H219A mutated UpVDAC proteins lost their metal binding ability, and became insensitive to metal-catalyzed oxidation (MCO) induced carbonylation. The overexpression of wild-type UpVDAC made yeast cells more sensitive to oxidative stress, retarded the growth of Arabidopsis seedlings and accelerated the seed aging, while overexpression of mutated UpVDAC weakened these effects of VDAC. These results reveal the relationship between the metal binding ability and carbonylation modification, as well as the probable function of VDAC in regulating cell vitality, seedling growth and seed aging.

Keywords: Ulmus pumila L.; metal-binding protein; metal-catalyzed oxidation (MCO); protein carbonylation; seed aging; voltage-dependent anion channel (VDAC).

Grants and funding

This work was supported by the National Natural Science Foundation of China (31971646, 91954202), and the National Training Program of Innovation and Entrepreneurship for Undergraduates (S202110022039).