Bridging the gap: investigating the role of phosphorylation at the serine 129 site of α-synuclein in VAPB-PTPIP51 interactions

Abstract Parkinson’s Disease (PD) is characterized by the aggregation and accumulation of α-synuclein (α-syn), along with abnormally high levels of α-syn phosphorylation at the serine 129 site (pSer 129 α-syn, p-α-syn). However, the mechanisms underlying the extensive phosphorylation at the serine 1...

Full description

Saved in:
Bibliographic Details
Main Authors: Weijin Liu, Yongquan Lu, Jia Liu, Yan Yu, Hui Yang
Format: Article
Language:English
Published: BMC 2025-02-01
Series:Acta Neuropathologica Communications
Subjects:
Online Access:https://doi.org/10.1186/s40478-025-01949-6
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849768249345966080
author Weijin Liu
Yongquan Lu
Jia Liu
Yan Yu
Hui Yang
author_facet Weijin Liu
Yongquan Lu
Jia Liu
Yan Yu
Hui Yang
author_sort Weijin Liu
collection DOAJ
description Abstract Parkinson’s Disease (PD) is characterized by the aggregation and accumulation of α-synuclein (α-syn), along with abnormally high levels of α-syn phosphorylation at the serine 129 site (pSer 129 α-syn, p-α-syn). However, the mechanisms underlying the extensive phosphorylation at the serine 129 site in the pathogenesis of PD, as well as the role of p-α-syn in the process, remain unclear. Furthermore, though α-syn could bind to VAPB and loosen Endoplasmic Reticulum (ER)-mitochondria associations by disrupting VAPB-PTPIP51 tethers, whether and how the phosphorylation of α-syn at the serine 129 site regulates VAPB-PTPIP51 interactions, remains unclear. Herein, Co-Immunoprecipitation and Mass Spectrometry (CO-IP/MS) studies were preformed to identify and compare the Protein-Protein Interactions (PPIs) of phosphorylated and total α-syn in the midbrains of Thy1-SNCA transgenic mice. We further performed CO-IP and Molecular Dynamics (MD) simulation assays to confirm the influence of α-syn phosphorylation on the aforementioned interactions. Additionally, we performed Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses to annotate the functional features of the common interacting proteins of p-α-syn and VAPB. The potential downstream proteins were further verified via CO-IP. According to the CO-IP and MD results, phosphorylation at the serine 129 site of α-syn increased VAPB-PTPIP51 interactions, and α-syn interacted directly with PTPIP51. Furthermore, functional and pathway enrichment analyses revealed that the common interacting proteins of p-α-syn and VAPB were significantly involved in protein binding, metal ion binding, structural constituent of the cytoskeleton, the intermediate filament cytoskeleton, and microtubule organization processes. Moreover, our findings confirmed the interactions of potential downstream target proteins (CLTC, CAMK2A, ATP1A3, and TUBB4B) with p-α-syn and VAPB. These findings collectively elucidate the structural underpinnings of serine 129 phosphorylation regulates the interaction between α-syn and both VAPB and PTPIP51. We hope that these findings will provide valuable insights into the role and regulatory mechanisms of serine 129 phosphorylation in the pathogenesis of pertinent diseases.
format Article
id doaj-art-66e5d28254b143b9bb3a3abbac6cc681
institution DOAJ
issn 2051-5960
language English
publishDate 2025-02-01
publisher BMC
record_format Article
series Acta Neuropathologica Communications
spelling doaj-art-66e5d28254b143b9bb3a3abbac6cc6812025-08-20T03:03:53ZengBMCActa Neuropathologica Communications2051-59602025-02-0113112110.1186/s40478-025-01949-6Bridging the gap: investigating the role of phosphorylation at the serine 129 site of α-synuclein in VAPB-PTPIP51 interactionsWeijin Liu0Yongquan Lu1Jia Liu2Yan Yu3Hui Yang4China Rehabilitation Science InstituteChongqing Key Laboratory of Emergency Medical, Chongqing Emergency Medical Center, Chongqing University Central HospitalBeijing Institute of Brain Disorders, Laboratory of Brain Disorders, Hypoxia Conditioning Translational Laboratory of Clinical Medicine, Capital Medical University, Collaborative Innovation Center for Brain Disorders, Beijing Advanced Innovation Center for Big Data-based Precision Medicine, Capital Medical UniversityChina Rehabilitation Science InstituteDepartment of Neurobiology, School of Basic Medical Sciences, Capital Medical UniversityAbstract Parkinson’s Disease (PD) is characterized by the aggregation and accumulation of α-synuclein (α-syn), along with abnormally high levels of α-syn phosphorylation at the serine 129 site (pSer 129 α-syn, p-α-syn). However, the mechanisms underlying the extensive phosphorylation at the serine 129 site in the pathogenesis of PD, as well as the role of p-α-syn in the process, remain unclear. Furthermore, though α-syn could bind to VAPB and loosen Endoplasmic Reticulum (ER)-mitochondria associations by disrupting VAPB-PTPIP51 tethers, whether and how the phosphorylation of α-syn at the serine 129 site regulates VAPB-PTPIP51 interactions, remains unclear. Herein, Co-Immunoprecipitation and Mass Spectrometry (CO-IP/MS) studies were preformed to identify and compare the Protein-Protein Interactions (PPIs) of phosphorylated and total α-syn in the midbrains of Thy1-SNCA transgenic mice. We further performed CO-IP and Molecular Dynamics (MD) simulation assays to confirm the influence of α-syn phosphorylation on the aforementioned interactions. Additionally, we performed Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses to annotate the functional features of the common interacting proteins of p-α-syn and VAPB. The potential downstream proteins were further verified via CO-IP. According to the CO-IP and MD results, phosphorylation at the serine 129 site of α-syn increased VAPB-PTPIP51 interactions, and α-syn interacted directly with PTPIP51. Furthermore, functional and pathway enrichment analyses revealed that the common interacting proteins of p-α-syn and VAPB were significantly involved in protein binding, metal ion binding, structural constituent of the cytoskeleton, the intermediate filament cytoskeleton, and microtubule organization processes. Moreover, our findings confirmed the interactions of potential downstream target proteins (CLTC, CAMK2A, ATP1A3, and TUBB4B) with p-α-syn and VAPB. These findings collectively elucidate the structural underpinnings of serine 129 phosphorylation regulates the interaction between α-syn and both VAPB and PTPIP51. We hope that these findings will provide valuable insights into the role and regulatory mechanisms of serine 129 phosphorylation in the pathogenesis of pertinent diseases.https://doi.org/10.1186/s40478-025-01949-6Parkinson’s diseaseα-synucleinSerine phosphorylationVAPBPTPIP51
spellingShingle Weijin Liu
Yongquan Lu
Jia Liu
Yan Yu
Hui Yang
Bridging the gap: investigating the role of phosphorylation at the serine 129 site of α-synuclein in VAPB-PTPIP51 interactions
Acta Neuropathologica Communications
Parkinson’s disease
α-synuclein
Serine phosphorylation
VAPB
PTPIP51
title Bridging the gap: investigating the role of phosphorylation at the serine 129 site of α-synuclein in VAPB-PTPIP51 interactions
title_full Bridging the gap: investigating the role of phosphorylation at the serine 129 site of α-synuclein in VAPB-PTPIP51 interactions
title_fullStr Bridging the gap: investigating the role of phosphorylation at the serine 129 site of α-synuclein in VAPB-PTPIP51 interactions
title_full_unstemmed Bridging the gap: investigating the role of phosphorylation at the serine 129 site of α-synuclein in VAPB-PTPIP51 interactions
title_short Bridging the gap: investigating the role of phosphorylation at the serine 129 site of α-synuclein in VAPB-PTPIP51 interactions
title_sort bridging the gap investigating the role of phosphorylation at the serine 129 site of α synuclein in vapb ptpip51 interactions
topic Parkinson’s disease
α-synuclein
Serine phosphorylation
VAPB
PTPIP51
url https://doi.org/10.1186/s40478-025-01949-6
work_keys_str_mv AT weijinliu bridgingthegapinvestigatingtheroleofphosphorylationattheserine129siteofasynucleininvapbptpip51interactions
AT yongquanlu bridgingthegapinvestigatingtheroleofphosphorylationattheserine129siteofasynucleininvapbptpip51interactions
AT jialiu bridgingthegapinvestigatingtheroleofphosphorylationattheserine129siteofasynucleininvapbptpip51interactions
AT yanyu bridgingthegapinvestigatingtheroleofphosphorylationattheserine129siteofasynucleininvapbptpip51interactions
AT huiyang bridgingthegapinvestigatingtheroleofphosphorylationattheserine129siteofasynucleininvapbptpip51interactions