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Enrichable cross-linkers for mapping direct protein interactions

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文献类型：外文期刊

作者： Wu, Ting ¹ ; Zhou, Hang-Xu ² ; Tian, Jing ⁴ ; Zhou, Rong ⁶ ; Huangfu, Shangwei ⁸ ; Jin, Bi-Kun ² ; Sablina, Anna ⁹ ; Zhou, Fangfang ¹ ; Chen, Hongli ⁸ ; Tang, Shibing ⁴ ; Zhang, Long ¹ ; Yang, Bing ² ;

作者机构： 1.Hangzhou City Univ, Sch Med, Key Lab Novel Targets & Drug Study Neural Repair Z, Hangzhou 310015, Zhejiang, Peoples R China

2.Zhejiang Univ, Affiliated Hosp 2, Life Sci Inst,Sch Med, Dept Med Oncol, Hangzhou 310058, Zhejiang, Peoples R China

3.Zhejiang Univ, Canc Ctr, Hangzhou 310058, Zhejiang, Peoples R China

4.Chinese Acad Sci, Inst Drug Discovery, Inst Biomed & Hlth, State Key Lab Resp Dis, Guangzhou 510530, Peoples R China

5.Chinese Acad Sci, Guangzhou Inst Biomed & Hlth, China New Zealand Belt & Rd Joint Lab Biomed & Hlt, Guangzhou 510530, Peoples R China

6.Chinese Acad Agr Sci, Inst Anim Sci, State Key Lab Anim Biotech Breeding, Beijing 100193, Peoples R China

7.Liaocheng Univ, Liaocheng 252059, Shandong, Peoples R China

8.ShanghaiTech Univ, Shanghai Inst Adv Immunochem Studies, Shanghai 201210, Peoples R China

9.Katholieke Univ Leuven, VIB, Ctr Canc Biol, B-3000 Louvain, Belgium

10.Soochow Univ, Inst Biol & Med Sci, Suzhou 215123, Jiangsu, Peoples R China

关键词： Cross-linking mass spectrometry; Thioredoxin; IMAC; Direct protein interaction; Subcellular organelles

期刊名称：GENOME BIOLOGY （影响因子：9.4；五年影响因子：16.3 ）

ISSN： 1474-760X

年卷期： 2025 年 26 卷 1 期

页码：

收录情况： SCI

摘要： BackgroundIt is crucial to investigate protein functions in specific subcellular environments. Cross-linking mass spectrometry is a powerful tool to map the direct interactome of proteins by identifying inter-protein cross-links. However, it is challenging to identify inter-protein cross-linked peptides due to their low abundance.ResultsWe chemically synthesize the cross-linkers ePDES1 and ePDES2 with an alkyne group and a compound with azide linked to a phosphate group to enrich for cross-linked peptides.ConclusionBased on the high-quality cross-linking spectra of ePDES1 and ePDES2, our methods achieve the identification of hundreds of directly interacting proteins or substrates of thioredoxins in the nucleus and mitochondria.

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Enrichable cross-linkers for mapping direct protein interactions

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