Spatiotemporal proteomics reveals the biosynthetic lysosomal membrane protein interactome in neurons

Abstract Lysosomes are membrane-bound organelles critical for maintaining cellular homeostasis. Delivery of biosynthetic lysosomal proteins to lysosomes is crucial to orchestrate proper lysosomal function. However, it remains unknown how the delivery of biosynthetic lysosomal proteins to lysosomes i...

Full description

Saved in:
Bibliographic Details
Main Authors: Chun Hei Li, Noortje Kersten, Nazmiye Özkan, Dan T. M. Nguyen, Max Koppers, Harm Post, Maarten Altelaar, Ginny G. Farias
Format: Article
Language:English
Published: Nature Portfolio 2024-12-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-024-55052-w
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849766499977265152
author Chun Hei Li
Noortje Kersten
Nazmiye Özkan
Dan T. M. Nguyen
Max Koppers
Harm Post
Maarten Altelaar
Ginny G. Farias
author_facet Chun Hei Li
Noortje Kersten
Nazmiye Özkan
Dan T. M. Nguyen
Max Koppers
Harm Post
Maarten Altelaar
Ginny G. Farias
author_sort Chun Hei Li
collection DOAJ
description Abstract Lysosomes are membrane-bound organelles critical for maintaining cellular homeostasis. Delivery of biosynthetic lysosomal proteins to lysosomes is crucial to orchestrate proper lysosomal function. However, it remains unknown how the delivery of biosynthetic lysosomal proteins to lysosomes is ensured in neurons, which are highly polarized cells. Here, we developed Protein Origin, Trafficking And Targeting to Organelle Mapping (POTATOMap), by combining trafficking synchronization and proximity-labelling based proteomics, to unravel the trafficking routes and interactome of the biosynthetic lysosomal membrane protein LAMP1 at specified time points. This approach, combined with advanced microscopy, enables us to identify the neuronal domain-specific trafficking machineries of biosynthetic LAMP1. We reveal a role in replenishing axonal lysosomes, in delivery of newly synthesized axonal synaptic proteins, and interactions with RNA granules to facilitate hitchhiking in the axon. POTATOMap offers a robust approach to map out dynamic biosynthetic protein trafficking and interactome from their origin to destination.
format Article
id doaj-art-0ccba86d7a0e4a858050ec3fde324a57
institution DOAJ
issn 2041-1723
language English
publishDate 2024-12-01
publisher Nature Portfolio
record_format Article
series Nature Communications
spelling doaj-art-0ccba86d7a0e4a858050ec3fde324a572025-08-20T03:04:34ZengNature PortfolioNature Communications2041-17232024-12-0115111910.1038/s41467-024-55052-wSpatiotemporal proteomics reveals the biosynthetic lysosomal membrane protein interactome in neuronsChun Hei Li0Noortje Kersten1Nazmiye Özkan2Dan T. M. Nguyen3Max Koppers4Harm Post5Maarten Altelaar6Ginny G. Farias7Cell Biology, Neurobiology and Biophysics, Department of Biology, Faculty of Science, Utrecht UniversityCell Biology, Neurobiology and Biophysics, Department of Biology, Faculty of Science, Utrecht UniversityCell Biology, Neurobiology and Biophysics, Department of Biology, Faculty of Science, Utrecht UniversityCell Biology, Neurobiology and Biophysics, Department of Biology, Faculty of Science, Utrecht UniversityCell Biology, Neurobiology and Biophysics, Department of Biology, Faculty of Science, Utrecht UniversityBiomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht UniversityBiomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht UniversityCell Biology, Neurobiology and Biophysics, Department of Biology, Faculty of Science, Utrecht UniversityAbstract Lysosomes are membrane-bound organelles critical for maintaining cellular homeostasis. Delivery of biosynthetic lysosomal proteins to lysosomes is crucial to orchestrate proper lysosomal function. However, it remains unknown how the delivery of biosynthetic lysosomal proteins to lysosomes is ensured in neurons, which are highly polarized cells. Here, we developed Protein Origin, Trafficking And Targeting to Organelle Mapping (POTATOMap), by combining trafficking synchronization and proximity-labelling based proteomics, to unravel the trafficking routes and interactome of the biosynthetic lysosomal membrane protein LAMP1 at specified time points. This approach, combined with advanced microscopy, enables us to identify the neuronal domain-specific trafficking machineries of biosynthetic LAMP1. We reveal a role in replenishing axonal lysosomes, in delivery of newly synthesized axonal synaptic proteins, and interactions with RNA granules to facilitate hitchhiking in the axon. POTATOMap offers a robust approach to map out dynamic biosynthetic protein trafficking and interactome from their origin to destination.https://doi.org/10.1038/s41467-024-55052-w
spellingShingle Chun Hei Li
Noortje Kersten
Nazmiye Özkan
Dan T. M. Nguyen
Max Koppers
Harm Post
Maarten Altelaar
Ginny G. Farias
Spatiotemporal proteomics reveals the biosynthetic lysosomal membrane protein interactome in neurons
Nature Communications
title Spatiotemporal proteomics reveals the biosynthetic lysosomal membrane protein interactome in neurons
title_full Spatiotemporal proteomics reveals the biosynthetic lysosomal membrane protein interactome in neurons
title_fullStr Spatiotemporal proteomics reveals the biosynthetic lysosomal membrane protein interactome in neurons
title_full_unstemmed Spatiotemporal proteomics reveals the biosynthetic lysosomal membrane protein interactome in neurons
title_short Spatiotemporal proteomics reveals the biosynthetic lysosomal membrane protein interactome in neurons
title_sort spatiotemporal proteomics reveals the biosynthetic lysosomal membrane protein interactome in neurons
url https://doi.org/10.1038/s41467-024-55052-w
work_keys_str_mv AT chunheili spatiotemporalproteomicsrevealsthebiosyntheticlysosomalmembraneproteininteractomeinneurons
AT noortjekersten spatiotemporalproteomicsrevealsthebiosyntheticlysosomalmembraneproteininteractomeinneurons
AT nazmiyeozkan spatiotemporalproteomicsrevealsthebiosyntheticlysosomalmembraneproteininteractomeinneurons
AT dantmnguyen spatiotemporalproteomicsrevealsthebiosyntheticlysosomalmembraneproteininteractomeinneurons
AT maxkoppers spatiotemporalproteomicsrevealsthebiosyntheticlysosomalmembraneproteininteractomeinneurons
AT harmpost spatiotemporalproteomicsrevealsthebiosyntheticlysosomalmembraneproteininteractomeinneurons
AT maartenaltelaar spatiotemporalproteomicsrevealsthebiosyntheticlysosomalmembraneproteininteractomeinneurons
AT ginnygfarias spatiotemporalproteomicsrevealsthebiosyntheticlysosomalmembraneproteininteractomeinneurons