The liprin-α/RIM complex regulates the dynamic assembly of presynaptic active zones via liquid-liquid phase separation.

Presynaptic scaffold proteins, including liprin-α, RIM, and ELKS, are pivotal to the assembly of the active zone and regulating the coupling of calcium signals and neurotransmitter release, yet the underlying mechanism remains poorly understood. Here, we determined the crystal structure of the lipri...

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Main Authors: Gaowei Jin, Joaquín Campos, Yang Liu, Berta Marcó de la Cruz, Shujing Zhang, Mingfu Liang, Kaiyue Li, Xingqiao Xie, Fredrik H Sterky, Claudio Acuna, Zhiyi Wei
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2025-06-01
Series:PLoS Biology
Online Access:https://doi.org/10.1371/journal.pbio.3002817
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author Gaowei Jin
Joaquín Campos
Yang Liu
Berta Marcó de la Cruz
Shujing Zhang
Mingfu Liang
Kaiyue Li
Xingqiao Xie
Fredrik H Sterky
Claudio Acuna
Zhiyi Wei
author_facet Gaowei Jin
Joaquín Campos
Yang Liu
Berta Marcó de la Cruz
Shujing Zhang
Mingfu Liang
Kaiyue Li
Xingqiao Xie
Fredrik H Sterky
Claudio Acuna
Zhiyi Wei
author_sort Gaowei Jin
collection DOAJ
description Presynaptic scaffold proteins, including liprin-α, RIM, and ELKS, are pivotal to the assembly of the active zone and regulating the coupling of calcium signals and neurotransmitter release, yet the underlying mechanism remains poorly understood. Here, we determined the crystal structure of the liprin-α2/RIM1 complex, revealing a multifaceted intermolecular interaction that drives the liprin-α/RIM assembly. Neurodevelopmental disease-associated mutations block the formation of the complex. Disrupting this interaction in cultured human neurons impairs synaptic transmission and reduces the readily releasable pool of synaptic vesicles. Super-resolution imaging analysis supports a role for liprin-α in recruiting RIM1 to the active zone, presumably by promoting the liquid-liquid phase separation (LLPS) of RIM1. Strikingly, the liprin-α/RIM interaction modulates the competitive distribution of ELKS1 and voltage-gated Ca2+ channels (VGCCs) in RIM1 condensates. Disrupting the liprin-α/RIM interaction significantly decreased VGCC accumulation in the condensed phase and rendered release more sensitive to the slow calcium buffer EGTA, suggesting an increased physical distance between VGCC and vesicular calcium sensors. Together, our findings provide a plausible mechanism of the liprin-α/RIM complex in regulating the coupling of calcium channels and primed synaptic vesicles via LLPS for efficient synaptic transmission and uncover the pathological implication of liprin-α mutations in neurodevelopmental disorders.
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spelling doaj-art-11a1e42d3cc141b1a67f588d533e974c2025-08-20T02:07:40ZengPublic Library of Science (PLoS)PLoS Biology1544-91731545-78852025-06-01236e300281710.1371/journal.pbio.3002817The liprin-α/RIM complex regulates the dynamic assembly of presynaptic active zones via liquid-liquid phase separation.Gaowei JinJoaquín CamposYang LiuBerta Marcó de la CruzShujing ZhangMingfu LiangKaiyue LiXingqiao XieFredrik H SterkyClaudio AcunaZhiyi WeiPresynaptic scaffold proteins, including liprin-α, RIM, and ELKS, are pivotal to the assembly of the active zone and regulating the coupling of calcium signals and neurotransmitter release, yet the underlying mechanism remains poorly understood. Here, we determined the crystal structure of the liprin-α2/RIM1 complex, revealing a multifaceted intermolecular interaction that drives the liprin-α/RIM assembly. Neurodevelopmental disease-associated mutations block the formation of the complex. Disrupting this interaction in cultured human neurons impairs synaptic transmission and reduces the readily releasable pool of synaptic vesicles. Super-resolution imaging analysis supports a role for liprin-α in recruiting RIM1 to the active zone, presumably by promoting the liquid-liquid phase separation (LLPS) of RIM1. Strikingly, the liprin-α/RIM interaction modulates the competitive distribution of ELKS1 and voltage-gated Ca2+ channels (VGCCs) in RIM1 condensates. Disrupting the liprin-α/RIM interaction significantly decreased VGCC accumulation in the condensed phase and rendered release more sensitive to the slow calcium buffer EGTA, suggesting an increased physical distance between VGCC and vesicular calcium sensors. Together, our findings provide a plausible mechanism of the liprin-α/RIM complex in regulating the coupling of calcium channels and primed synaptic vesicles via LLPS for efficient synaptic transmission and uncover the pathological implication of liprin-α mutations in neurodevelopmental disorders.https://doi.org/10.1371/journal.pbio.3002817
spellingShingle Gaowei Jin
Joaquín Campos
Yang Liu
Berta Marcó de la Cruz
Shujing Zhang
Mingfu Liang
Kaiyue Li
Xingqiao Xie
Fredrik H Sterky
Claudio Acuna
Zhiyi Wei
The liprin-α/RIM complex regulates the dynamic assembly of presynaptic active zones via liquid-liquid phase separation.
PLoS Biology
title The liprin-α/RIM complex regulates the dynamic assembly of presynaptic active zones via liquid-liquid phase separation.
title_full The liprin-α/RIM complex regulates the dynamic assembly of presynaptic active zones via liquid-liquid phase separation.
title_fullStr The liprin-α/RIM complex regulates the dynamic assembly of presynaptic active zones via liquid-liquid phase separation.
title_full_unstemmed The liprin-α/RIM complex regulates the dynamic assembly of presynaptic active zones via liquid-liquid phase separation.
title_short The liprin-α/RIM complex regulates the dynamic assembly of presynaptic active zones via liquid-liquid phase separation.
title_sort liprin α rim complex regulates the dynamic assembly of presynaptic active zones via liquid liquid phase separation
url https://doi.org/10.1371/journal.pbio.3002817
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