Global Analysis of Protein and Small-Molecule Substrates of Ubiquitin-Like Proteins

doi:10.1016/j.mcpro.2025.100975

	Global Analysis of Protein and Small-Molecule Substrates of Ubiquitin-Like Proteins
	Shao, Guang-Can 1; Chen, Zhen-Lin 2,3; Lu, Shan 1; Wu, Qing-Cui 1; Sheng, Yao 1; Wang, Jing 1; Ma, Yan 1,4; Sui, Jian-Hua 1,4; Chi, Hao 2,3; Qi, Xiang-Bing 1,4; He, Si-Min 1,4; Du, Li-Lin 1,4; Dong, Meng-Qiu 1,4
	2025-07-01
发表期刊	MOLECULAR & CELLULAR PROTEOMICS
卷号	24 期号:7 页码:20
摘要	Ubiquitin-like proteins (UBLs) constitute a family of evolutionarily conserved proteins that share similarities with ubiquitin in 3D structures and modification mechanisms. For most UBLs including small-ubiquitin-like modifiers (SUMO), their modification sites on substrate proteins cannot be identified using the mass spectrometry-based method that has been successful for identifying ubiquitination sites, unless a UBL protein is mutated accordingly. To identify UBL modification sites without having to mutate UBL, we have developed a dedicated search engine pLink-UBL on the basis of pLink, a software tool for the identification of cross-linked peptide pairs. pLink-UBL exhibited superior precision, sensitivity, and speed than "make-do" search engines such as MaxQuant, pFind, and pLink. For example, compared to MaxQuant, pLink-UBL increased the number of identified SUMOylation sites by 50 similar to 300% from the same datasets. Additionally, we present a method for identifying small molecule modifications of UBLs. This method involves antibody enrichment of a UBL C-terminal peptide following enrichment of a UBL protein, followed by LC-MS/MS analysis and a pFind 3 blind search to identify unexpected modifications. Using this method, we have discovered nonprotein substrates of SUMO, of which spermidine is the major one for fission yeast SUMO Pmt3. Spermidine can be conjugated to the C-terminal carboxylate group of Pmt3 through its N1 or also likely, N8 amino group in the presence of SUMO E1, E2, and ATP. Pmt3-spermidine conjugation does not require E3 and can be reversed by SUMO isopeptidase Ulp1. SUMO-spermidine conjugation is present in mice and humans. Also, spermidine can be conjugated to ubiquitin in vitro by E1 and E2 in the presence of ATP. The above observations suggest that spermidine may be a common small molecule substrate of SUMO and possibly ubiquitin across eukaryotic species.
DOI	10.1016/j.mcpro.2025.100975
收录类别	SCI
语种	英语
资助项目	NIBS, Beijing ; TIMBR, Tsinghua University ; National Natural Science Foundation[2167050459]
WOS研究方向	Biochemistry & Molecular Biology
WOS类目	Biochemical Research Methods
WOS记录号	WOS:001527077100002
出版者	ELSEVIER
引用统计
文献类型	期刊论文
条目标识符	http://119.78.100.204/handle/2XEOYT63/42057
专题	中国科学院计算技术研究所期刊论文_英文
通讯作者	He, Si-Min; Du, Li-Lin; Dong, Meng-Qiu
作者单位	1.Natl Inst Biol Sci, Beijing, Peoples R China 2.Chinese Acad Sci, Inst Comp Technol, Key Lab Intelligent Informat Proc, Beijing, Peoples R China 3.Univ Chinese Acad Sci, Beijing, Peoples R China 4.Tsinghua Univ, Tsinghua Inst Multidisciplinary Biomed Res, Beijing, Peoples R China
推荐引用方式 GB/T 7714	Shao, Guang-Can,Chen, Zhen-Lin,Lu, Shan,et al. Global Analysis of Protein and Small-Molecule Substrates of Ubiquitin-Like Proteins[J]. MOLECULAR & CELLULAR PROTEOMICS,2025,24(7):20.
APA	Shao, Guang-Can.,Chen, Zhen-Lin.,Lu, Shan.,Wu, Qing-Cui.,Sheng, Yao.,...&Dong, Meng-Qiu.(2025).Global Analysis of Protein and Small-Molecule Substrates of Ubiquitin-Like Proteins.MOLECULAR & CELLULAR PROTEOMICS,24(7),20.
MLA	Shao, Guang-Can,et al."Global Analysis of Protein and Small-Molecule Substrates of Ubiquitin-Like Proteins".MOLECULAR & CELLULAR PROTEOMICS 24.7(2025):20.