O-GlcNAcylation modulates liquid–liquid phase separation of SynGAP/PSD-95

Lv，Pinou1,2,3; Du，Yifei1,2,3; He，Changdong4,5; Peng，Luxin1,3; Zhou，Xinyue1,2,3; Wan，Yi1,2,3; Zeng，Menglong6; Zhou，Wen1,3; Zou，Peng1,2,3,7,8; Li，Chenjian2,9; Zhang，Mingjie6,10; Dong，Suwei4,5; Chen，Xing1,2,3,7,8

2022

10.1038/s41557-022-00946-9

Nature Chemistry   影响因子和分区

1755-4330

1755-4349

Liquid–liquid phase separation (LLPS) of SynGAP and PSD-95, two abundant proteins that interact in the postsynaptic density (PSD) of neurons, has been implicated in modulating SynGAP PSD enrichment in excitatory synapses. However, the underlying regulatory mechanisms remain enigmatic. Here we report that O-GlcNAcylation of SynGAP acts as a suppressor of LLPS of the SynGAP/PSD-95 complex. We identified multiple O-GlcNAc modification sites for the endogenous SynGAP isolated from rat brain and the recombinantly expressed protein. Protein semisynthesis was used to generate site-specifically O-GlcNAcylated forms of SynGAP, and in vitro and cell-based LLPS assays demonstrated that T1306 O-GlcNAc of SynGAP blocks the interaction with PSD-95, thus inhibiting LLPS. Furthermore, O-GlcNAcylation suppresses SynGAP/PSD-95 LLPS in a dominant-negative manner, enabling sub-stoichiometric O-GlcNAcylation to exert effective regulation. We also showed that O-GlcNAc-dependent LLPS is reversibly regulated by O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA). These findings demonstrate that OGT- and OGA-catalysed O-GlcNAc cycling may serve as an LLPS-regulating post-translational modification. [Figure not available: see fulltext.]

SCI

英语

NI论文

其他

WOS:000802830200001

2-s2.0-85131052947

Scopus

1.College of Chemistry and Molecular Engineering,Peking University,Beijing,China
2.Peking-Tsinghua Center for Life Sciences,Peking University,Beijing,China
3.Beijing National Laboratory for Molecular Sciences,Peking University,Beijing,China
4.State Key Laboratory of Natural and Biomimetic Drugs,Peking University,Beijing,China
5.School of Pharmaceutical Sciences,Peking University,Beijing,China
6.Division of Life Science,State Key Laboratory of Molecular Neuroscience,Hong Kong University of Science and Technology,Hong Kong
7.Synthetic and Functional Biomolecules Center,Peking University,Beijing,China
8.Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education,Peking University,Beijing,China
9.School of Life Sciences,Peking University,Beijing,China
10.School of Life Sciences,Southern University of Science and Technology,Shenzhen,China

Lv，Pinou,Du，Yifei,He，Changdong,等. O-GlcNAcylation modulates liquid–liquid phase separation of SynGAP/PSD-95[J]. Nature Chemistry,2022.

Lv，Pinou.,Du，Yifei.,He，Changdong.,Peng，Luxin.,Zhou，Xinyue.,...&Chen，Xing.(2022).O-GlcNAcylation modulates liquid–liquid phase separation of SynGAP/PSD-95.Nature Chemistry.

Lv，Pinou,et al."O-GlcNAcylation modulates liquid–liquid phase separation of SynGAP/PSD-95".Nature Chemistry (2022).