In Situ Peroxidase Labeling and Mass-Spectrometry Connects Alpha-Synuclein Directly to Endocytic Trafficking and mRNA Metabolism in Neurons

Chee Yeun Chung; Vikram Khurana; Song Yi; Nidhi Sahni; Ken H. Loh; Pavan K. Auluck; Valeriya Baru; Namrata D. Udeshi; Yelena Freyzon; Steven A. Carr; David E. Hill; Marc Vidal; Alice Y. Ting; Susan Lindquist

doi:10.1016/j.cels.2017.01.002

In Situ Peroxidase Labeling and Mass-Spectrometry Connects Alpha-Synuclein Directly to Endocytic Trafficking and mRNA Metabolism in Neurons

Chee Yeun Chung, Vikram Khurana, Song Yi, Nidhi Sahni, Ken H. Loh, Pavan K. Auluck, Valeriya Baru, Namrata D. Udeshi, Yelena Freyzon, Steven A. Carr, David E. Hill, Marc Vidal, Alice Y. Ting, Susan Lindquist

Systems Biology

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83 Scopus citations

Abstract

Synucleinopathies, including Parkinson's disease (PD), are associated with the misfolding and mistrafficking of alpha-synuclein (α-syn). Here, using an ascorbate peroxidase (APEX)-based labeling method combined with mass spectrometry, we defined a network of proteins in the immediate vicinity of α-syn in living neurons to shed light on α-syn function. This approach identified 225 proteins, including synaptic proteins, proteins involved in endocytic vesicle trafficking, the retromer complex, phosphatases and mRNA binding proteins. Many were in complexes with α-syn, and some were encoded by genes known to be risk factors for PD and other neurodegenerative diseases. Endocytic trafficking and mRNA translation proteins within this spatial α-syn map overlapped with genetic modifiers of α-syn toxicity, developed in an accompanying study (Khurana et al., this issue of Cell Systems). Our data suggest that perturbation of these particular pathways is directly related to the spatial localization of α-syn within the cell. These approaches provide new avenues to systematically examine protein function and pathology in living cells.

Original language	English (US)
Pages (from-to)	242-250.e4
Journal	Cell Systems
Volume	4
Issue number	2
DOIs	https://doi.org/10.1016/j.cels.2017.01.002
State	Published - Feb 22 2017

ASJC Scopus subject areas

Pathology and Forensic Medicine
Histology
Cell Biology

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Chung, C. Y., Khurana, V., Yi, S., Sahni, N., Loh, K. H., Auluck, P. K., Baru, V., Udeshi, N. D., Freyzon, Y., Carr, S. A., Hill, D. E., Vidal, M., Ting, A. Y., & Lindquist, S. (2017). In Situ Peroxidase Labeling and Mass-Spectrometry Connects Alpha-Synuclein Directly to Endocytic Trafficking and mRNA Metabolism in Neurons. Cell Systems, 4(2), 242-250.e4. https://doi.org/10.1016/j.cels.2017.01.002

Chung, CY, Khurana, V, Yi, S, Sahni, N, Loh, KH, Auluck, PK, Baru, V, Udeshi, ND, Freyzon, Y, Carr, SA, Hill, DE, Vidal, M, Ting, AY & Lindquist, S 2017, 'In Situ Peroxidase Labeling and Mass-Spectrometry Connects Alpha-Synuclein Directly to Endocytic Trafficking and mRNA Metabolism in Neurons', Cell Systems, vol. 4, no. 2, pp. 242-250.e4. https://doi.org/10.1016/j.cels.2017.01.002

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abstract = "Synucleinopathies, including Parkinson's disease (PD), are associated with the misfolding and mistrafficking of alpha-synuclein (α-syn). Here, using an ascorbate peroxidase (APEX)-based labeling method combined with mass spectrometry, we defined a network of proteins in the immediate vicinity of α-syn in living neurons to shed light on α-syn function. This approach identified 225 proteins, including synaptic proteins, proteins involved in endocytic vesicle trafficking, the retromer complex, phosphatases and mRNA binding proteins. Many were in complexes with α-syn, and some were encoded by genes known to be risk factors for PD and other neurodegenerative diseases. Endocytic trafficking and mRNA translation proteins within this spatial α-syn map overlapped with genetic modifiers of α-syn toxicity, developed in an accompanying study (Khurana et al., this issue of Cell Systems). Our data suggest that perturbation of these particular pathways is directly related to the spatial localization of α-syn within the cell. These approaches provide new avenues to systematically examine protein function and pathology in living cells.",

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AU - Freyzon, Yelena

AU - Carr, Steven A.

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N2 - Synucleinopathies, including Parkinson's disease (PD), are associated with the misfolding and mistrafficking of alpha-synuclein (α-syn). Here, using an ascorbate peroxidase (APEX)-based labeling method combined with mass spectrometry, we defined a network of proteins in the immediate vicinity of α-syn in living neurons to shed light on α-syn function. This approach identified 225 proteins, including synaptic proteins, proteins involved in endocytic vesicle trafficking, the retromer complex, phosphatases and mRNA binding proteins. Many were in complexes with α-syn, and some were encoded by genes known to be risk factors for PD and other neurodegenerative diseases. Endocytic trafficking and mRNA translation proteins within this spatial α-syn map overlapped with genetic modifiers of α-syn toxicity, developed in an accompanying study (Khurana et al., this issue of Cell Systems). Our data suggest that perturbation of these particular pathways is directly related to the spatial localization of α-syn within the cell. These approaches provide new avenues to systematically examine protein function and pathology in living cells.

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In Situ Peroxidase Labeling and Mass-Spectrometry Connects Alpha-Synuclein Directly to Endocytic Trafficking and mRNA Metabolism in Neurons

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