Application of Reducible Covalent Capture Purification for Resolving Persulfidome and Nucleolin S-Sulfhydration

被引：0

作者：

Huang, Wei-Chieh ^{[1
]}

Hsu, Kai-Wen ^{[2
,3
,4
]}

Peng, Pei-Hua ^{[2
]}

Zeng, Wan-Ting ^{[5
]}

Gu, Ting-Jia ^{[5
]}

Lin, Li-Jie ^{[2
,3
,6
]}

Hsieh, Min-Tsang ^{[7
,8
,9
]}

Lee, Der-Yen ^{[5
]}

Chang, Geen-Dong ^{[1
]}

机构：

[1] Natl Taiwan Univ, Grad Inst Biochem Sci, Taipei 106, Taiwan

[2] China Med Univ, Drug Dev Ctr, Program Canc Biol & Drug Discovery, Taichung 40402, Taiwan

[3] China Med Univ, Res Ctr Canc Biol, Taichung 40402, Taiwan

[4] China Med Univ, Inst Translat Med & New Drug Dev, Taichung 40402, Taiwan

[5] China Med Univ, Grad Inst Integrated Med, Taichung 40402, Taiwan

[6] China Med Univ, Acad Sinica, Ph D Program Canc Biol & Drug Discovery, Taichung 40402, Taiwan

[7] China Med Univ, Sch Pharm, Taichung, Taiwan

[8] China Med Univ, Drug Dev Ctr, Taichung, Taiwan

[9] China Med Univ Hosp, Chinese Med Res & Dev Ctr, Taichung 40447, Taiwan

来源：

ANALYTICAL CHEMISTRY | 2024年 / 96卷 / 35期

关键词：

RIBOSOMAL-PROTEIN L26; BCL-2; MESSENGER-RNA; DNA-DAMAGE; TRANSLATION;

D O I：

10.1021/acs.analchem.4c02717

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Protein S-sulfhydration involves the regulation of various protein functions, and resolving the S-sulfhydrated proteome (persulfidome) allows for a deeper exploration of various redox regulations. Therefore, we designed a reducible covalent capture method for isolating S-sulfhydrated proteins, which can analyze the persulfidome in biological samples and monitor specific S-sulfhydrated proteins. In this study, we applied this method to reveal the S-sulfhydration levels of proteins, including 3-phosphoglyceraldehyde dehydrogenase, NF kappa B/p65, and nucleolin. Furthermore, this technique can be used to enrich S-sulfhydrated peptides, aiding in the determination of protein S-sulfhydration modification sites. Finally, we observed that the S-sulfhydration and oxidation of nucleolin on the C543 residue correlate with its nuclear translocation, downstream regulation of p53, Bcl-xL, and Bcl-2 RNA levels and protein expression, as well as the protective function against oxidative stress. Therefore, this method may facilitate the understanding of the regulation of protein function by redox perturbation.

引用

页码：14186 / 14196

页数：11