Targeted Analysis of Mitochondrial Protein Conformations and Interactions by Endogenous ROS‐Triggered Cross‐Linker Release
Abstract The study of in situ conformations and interactions of mitochondrial proteins plays a crucial role in understanding their biological functions. Current chemical cross‐linking mass spectrometry (CX‐MS) has difficulty in achieving in‐depth analysis of mitochondrial proteins for cells without...
Saved in:
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2024-12-01
|
| Series: | Advanced Science |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/advs.202408462 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1846106583237394432 |
|---|---|
| author | Wen Zhou Yuwan Chen Wenxin Fu Xinwei Li Yufei Xia Qun Zhao Baofeng Zhao Yukui Zhang Kaiguang Yang Lihua Zhang |
| author_facet | Wen Zhou Yuwan Chen Wenxin Fu Xinwei Li Yufei Xia Qun Zhao Baofeng Zhao Yukui Zhang Kaiguang Yang Lihua Zhang |
| author_sort | Wen Zhou |
| collection | DOAJ |
| description | Abstract The study of in situ conformations and interactions of mitochondrial proteins plays a crucial role in understanding their biological functions. Current chemical cross‐linking mass spectrometry (CX‐MS) has difficulty in achieving in‐depth analysis of mitochondrial proteins for cells without genetic modification. Herein, this work develops the reactive oxygen species (ROS)‐responsive cross‐linker delivery nanoparticles (R‐CDNP) targeting mitochondria. R‐CDNP contains mitochondria‐targeting module triphenylphosphine, ROS‐responsive module thioketal, loading module poly(lactic‐co‐glycolic acid) (PLGA), and polyethylene glycol (PEG), and cross‐linker module disuccinimidyl suberate (DSS). After targeting mitochondria, ROS‐triggered cross‐linker release improves the cross‐linking coverage of mitochondria in situ. In total, this work identifies 2103 cross‐linked sites of 572 mitochondrial proteins in HepG2 cells. 1718 intra‐links reveal dynamic conformations involving chaperones with ATP‐dependent conformation cycles, and 385 inter‐links reveal dynamic interactions involving OXPHOS complexes and 27 pairs of possible potential interactions. These results signify that R‐CDNP can achieve dynamic conformation and interaction analysis of mitochondrial proteins in living cells, thereby contributing to a better understanding of their biological functions. |
| format | Article |
| id | doaj-art-a263694dba404206a4aa908a0382a3b3 |
| institution | Kabale University |
| issn | 2198-3844 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Wiley |
| record_format | Article |
| series | Advanced Science |
| spelling | doaj-art-a263694dba404206a4aa908a0382a3b32024-12-27T13:00:46ZengWileyAdvanced Science2198-38442024-12-011148n/an/a10.1002/advs.202408462Targeted Analysis of Mitochondrial Protein Conformations and Interactions by Endogenous ROS‐Triggered Cross‐Linker ReleaseWen Zhou0Yuwan Chen1Wenxin Fu2Xinwei Li3Yufei Xia4Qun Zhao5Baofeng Zhao6Yukui Zhang7Kaiguang Yang8Lihua Zhang9State Key Laboratory of Medical Proteomics National Chromatographic R. & A. Center CAS Key Laboratory of Separation Science for Analytical Chemistry Dalian Institute of Chemical Physics Chinese Academy of Sciences 457 Zhongshan Road Dalian 116023 ChinaState Key Laboratory of Medical Proteomics National Chromatographic R. & A. Center CAS Key Laboratory of Separation Science for Analytical Chemistry Dalian Institute of Chemical Physics Chinese Academy of Sciences 457 Zhongshan Road Dalian 116023 ChinaState Key Laboratory of Medical Proteomics National Chromatographic R. & A. Center CAS Key Laboratory of Separation Science for Analytical Chemistry Dalian Institute of Chemical Physics Chinese Academy of Sciences 457 Zhongshan Road Dalian 116023 ChinaState Key Laboratory of Medical Proteomics National Chromatographic R. & A. Center CAS Key Laboratory of Separation Science for Analytical Chemistry Dalian Institute of Chemical Physics Chinese Academy of Sciences 457 Zhongshan Road Dalian 116023 ChinaUniversity of Chinese Academy of Sciences Beijing 100049 ChinaState Key Laboratory of Medical Proteomics National Chromatographic R. & A. Center CAS Key Laboratory of Separation Science for Analytical Chemistry Dalian Institute of Chemical Physics Chinese Academy of Sciences 457 Zhongshan Road Dalian 116023 ChinaState Key Laboratory of Medical Proteomics National Chromatographic R. & A. Center CAS Key Laboratory of Separation Science for Analytical Chemistry Dalian Institute of Chemical Physics Chinese Academy of Sciences 457 Zhongshan Road Dalian 116023 ChinaState Key Laboratory of Medical Proteomics National Chromatographic R. & A. Center CAS Key Laboratory of Separation Science for Analytical Chemistry Dalian Institute of Chemical Physics Chinese Academy of Sciences 457 Zhongshan Road Dalian 116023 ChinaState Key Laboratory of Medical Proteomics National Chromatographic R. & A. Center CAS Key Laboratory of Separation Science for Analytical Chemistry Dalian Institute of Chemical Physics Chinese Academy of Sciences 457 Zhongshan Road Dalian 116023 ChinaState Key Laboratory of Medical Proteomics National Chromatographic R. & A. Center CAS Key Laboratory of Separation Science for Analytical Chemistry Dalian Institute of Chemical Physics Chinese Academy of Sciences 457 Zhongshan Road Dalian 116023 ChinaAbstract The study of in situ conformations and interactions of mitochondrial proteins plays a crucial role in understanding their biological functions. Current chemical cross‐linking mass spectrometry (CX‐MS) has difficulty in achieving in‐depth analysis of mitochondrial proteins for cells without genetic modification. Herein, this work develops the reactive oxygen species (ROS)‐responsive cross‐linker delivery nanoparticles (R‐CDNP) targeting mitochondria. R‐CDNP contains mitochondria‐targeting module triphenylphosphine, ROS‐responsive module thioketal, loading module poly(lactic‐co‐glycolic acid) (PLGA), and polyethylene glycol (PEG), and cross‐linker module disuccinimidyl suberate (DSS). After targeting mitochondria, ROS‐triggered cross‐linker release improves the cross‐linking coverage of mitochondria in situ. In total, this work identifies 2103 cross‐linked sites of 572 mitochondrial proteins in HepG2 cells. 1718 intra‐links reveal dynamic conformations involving chaperones with ATP‐dependent conformation cycles, and 385 inter‐links reveal dynamic interactions involving OXPHOS complexes and 27 pairs of possible potential interactions. These results signify that R‐CDNP can achieve dynamic conformation and interaction analysis of mitochondrial proteins in living cells, thereby contributing to a better understanding of their biological functions.https://doi.org/10.1002/advs.202408462cross‐linking mass spectrometrymitochondriananoparticlesprotein complexesreactive oxygen species‐responsive |
| spellingShingle | Wen Zhou Yuwan Chen Wenxin Fu Xinwei Li Yufei Xia Qun Zhao Baofeng Zhao Yukui Zhang Kaiguang Yang Lihua Zhang Targeted Analysis of Mitochondrial Protein Conformations and Interactions by Endogenous ROS‐Triggered Cross‐Linker Release Advanced Science cross‐linking mass spectrometry mitochondria nanoparticles protein complexes reactive oxygen species‐responsive |
| title | Targeted Analysis of Mitochondrial Protein Conformations and Interactions by Endogenous ROS‐Triggered Cross‐Linker Release |
| title_full | Targeted Analysis of Mitochondrial Protein Conformations and Interactions by Endogenous ROS‐Triggered Cross‐Linker Release |
| title_fullStr | Targeted Analysis of Mitochondrial Protein Conformations and Interactions by Endogenous ROS‐Triggered Cross‐Linker Release |
| title_full_unstemmed | Targeted Analysis of Mitochondrial Protein Conformations and Interactions by Endogenous ROS‐Triggered Cross‐Linker Release |
| title_short | Targeted Analysis of Mitochondrial Protein Conformations and Interactions by Endogenous ROS‐Triggered Cross‐Linker Release |
| title_sort | targeted analysis of mitochondrial protein conformations and interactions by endogenous ros triggered cross linker release |
| topic | cross‐linking mass spectrometry mitochondria nanoparticles protein complexes reactive oxygen species‐responsive |
| url | https://doi.org/10.1002/advs.202408462 |
| work_keys_str_mv | AT wenzhou targetedanalysisofmitochondrialproteinconformationsandinteractionsbyendogenousrostriggeredcrosslinkerrelease AT yuwanchen targetedanalysisofmitochondrialproteinconformationsandinteractionsbyendogenousrostriggeredcrosslinkerrelease AT wenxinfu targetedanalysisofmitochondrialproteinconformationsandinteractionsbyendogenousrostriggeredcrosslinkerrelease AT xinweili targetedanalysisofmitochondrialproteinconformationsandinteractionsbyendogenousrostriggeredcrosslinkerrelease AT yufeixia targetedanalysisofmitochondrialproteinconformationsandinteractionsbyendogenousrostriggeredcrosslinkerrelease AT qunzhao targetedanalysisofmitochondrialproteinconformationsandinteractionsbyendogenousrostriggeredcrosslinkerrelease AT baofengzhao targetedanalysisofmitochondrialproteinconformationsandinteractionsbyendogenousrostriggeredcrosslinkerrelease AT yukuizhang targetedanalysisofmitochondrialproteinconformationsandinteractionsbyendogenousrostriggeredcrosslinkerrelease AT kaiguangyang targetedanalysisofmitochondrialproteinconformationsandinteractionsbyendogenousrostriggeredcrosslinkerrelease AT lihuazhang targetedanalysisofmitochondrialproteinconformationsandinteractionsbyendogenousrostriggeredcrosslinkerrelease |