Cryo-TEM structure of β-glucocerebrosidase in complex with its transporter LIMP-2

Abstract Targeting proteins to their final cellular destination requires transport mechanisms and nearly all lysosomal enzymes reach the lysosome via the mannose-6-phosphate receptor pathway. One of the few known exceptions is the enzyme β-glucocerebrosidase (GCase) that requires the lysosomal integ...

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Main Authors: Jan Philipp Dobert, Jan-Hannes Schäfer, Thomas Dal Maso, Priyadarshini Ravindran, Dustin J. E. Huard, Eileen Socher, Lisa A. Schildmeyer, Raquel L. Lieberman, Wim Versées, Arne Moeller, Friederike Zunke, Philipp Arnold
Format: Article
Language:English
Published: Nature Portfolio 2025-03-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-58340-1
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author Jan Philipp Dobert
Jan-Hannes Schäfer
Thomas Dal Maso
Priyadarshini Ravindran
Dustin J. E. Huard
Eileen Socher
Lisa A. Schildmeyer
Raquel L. Lieberman
Wim Versées
Arne Moeller
Friederike Zunke
Philipp Arnold
author_facet Jan Philipp Dobert
Jan-Hannes Schäfer
Thomas Dal Maso
Priyadarshini Ravindran
Dustin J. E. Huard
Eileen Socher
Lisa A. Schildmeyer
Raquel L. Lieberman
Wim Versées
Arne Moeller
Friederike Zunke
Philipp Arnold
author_sort Jan Philipp Dobert
collection DOAJ
description Abstract Targeting proteins to their final cellular destination requires transport mechanisms and nearly all lysosomal enzymes reach the lysosome via the mannose-6-phosphate receptor pathway. One of the few known exceptions is the enzyme β-glucocerebrosidase (GCase) that requires the lysosomal integral membrane protein type-2 (LIMP-2) as a proprietary lysosomal transporter. Genetic variations in the GCase encoding gene GBA1 cause Gaucher’s disease (GD) and present the highest genetic risk factor to develop Parkinson’s disease (PD). Activators targeting GCase emerge as a promising therapeutic approach to treat GD and PD, with pre-clinical and clinical trials ongoing. In this study, we resolve the complex of GCase and LIMP-2 using cryo-electron microscopy with the aid of an engineered LIMP-2 shuttle and two GCase-targeted pro-macrobodies. We identify helix 5 and helix 7 of LIMP-2 to interact with a binding pocket in GCase, forming a mostly hydrophobic interaction interface supported by one essential salt bridge. Understanding the interplay of GCase and LIMP-2 on a structural level is crucial to identify potential activation sites and conceptualizing novel therapeutic approaches targeting GCase. Here, we unveil the protein structure of a mannose-6-phosphate-independent lysosomal transport complex and provide fundamental knowledge for translational clinical research to overcome GD and PD.
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spelling doaj-art-3d75a6e016a840db84145ab0d9b4dfe22025-08-20T02:10:21ZengNature PortfolioNature Communications2041-17232025-03-0116111410.1038/s41467-025-58340-1Cryo-TEM structure of β-glucocerebrosidase in complex with its transporter LIMP-2Jan Philipp Dobert0Jan-Hannes Schäfer1Thomas Dal Maso2Priyadarshini Ravindran3Dustin J. E. Huard4Eileen Socher5Lisa A. Schildmeyer6Raquel L. Lieberman7Wim Versées8Arne Moeller9Friederike Zunke10Philipp Arnold11Department of Molecular Neurology, University Hospital Erlangen, Friedrich-Alexander-University Erlangen-Nürnberg (FAU)Department of Biology/Chemistry, Structural Biology Section, Osnabrück UniversityVIB-VUB Center for Structural Biology, VIB, Pleinlaan 2Department of Molecular Neurology, University Hospital Erlangen, Friedrich-Alexander-University Erlangen-Nürnberg (FAU)School of Chemistry and Biochemistry, Georgia Institute of TechnologyInstitute of Functional and Clinical Anatomy, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU)School of Chemistry and Biochemistry, Georgia Institute of TechnologySchool of Chemistry and Biochemistry, Georgia Institute of TechnologyVIB-VUB Center for Structural Biology, VIB, Pleinlaan 2Department of Biology/Chemistry, Structural Biology Section, Osnabrück UniversityDepartment of Molecular Neurology, University Hospital Erlangen, Friedrich-Alexander-University Erlangen-Nürnberg (FAU)Institute of Functional and Clinical Anatomy, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU)Abstract Targeting proteins to their final cellular destination requires transport mechanisms and nearly all lysosomal enzymes reach the lysosome via the mannose-6-phosphate receptor pathway. One of the few known exceptions is the enzyme β-glucocerebrosidase (GCase) that requires the lysosomal integral membrane protein type-2 (LIMP-2) as a proprietary lysosomal transporter. Genetic variations in the GCase encoding gene GBA1 cause Gaucher’s disease (GD) and present the highest genetic risk factor to develop Parkinson’s disease (PD). Activators targeting GCase emerge as a promising therapeutic approach to treat GD and PD, with pre-clinical and clinical trials ongoing. In this study, we resolve the complex of GCase and LIMP-2 using cryo-electron microscopy with the aid of an engineered LIMP-2 shuttle and two GCase-targeted pro-macrobodies. We identify helix 5 and helix 7 of LIMP-2 to interact with a binding pocket in GCase, forming a mostly hydrophobic interaction interface supported by one essential salt bridge. Understanding the interplay of GCase and LIMP-2 on a structural level is crucial to identify potential activation sites and conceptualizing novel therapeutic approaches targeting GCase. Here, we unveil the protein structure of a mannose-6-phosphate-independent lysosomal transport complex and provide fundamental knowledge for translational clinical research to overcome GD and PD.https://doi.org/10.1038/s41467-025-58340-1
spellingShingle Jan Philipp Dobert
Jan-Hannes Schäfer
Thomas Dal Maso
Priyadarshini Ravindran
Dustin J. E. Huard
Eileen Socher
Lisa A. Schildmeyer
Raquel L. Lieberman
Wim Versées
Arne Moeller
Friederike Zunke
Philipp Arnold
Cryo-TEM structure of β-glucocerebrosidase in complex with its transporter LIMP-2
Nature Communications
title Cryo-TEM structure of β-glucocerebrosidase in complex with its transporter LIMP-2
title_full Cryo-TEM structure of β-glucocerebrosidase in complex with its transporter LIMP-2
title_fullStr Cryo-TEM structure of β-glucocerebrosidase in complex with its transporter LIMP-2
title_full_unstemmed Cryo-TEM structure of β-glucocerebrosidase in complex with its transporter LIMP-2
title_short Cryo-TEM structure of β-glucocerebrosidase in complex with its transporter LIMP-2
title_sort cryo tem structure of β glucocerebrosidase in complex with its transporter limp 2
url https://doi.org/10.1038/s41467-025-58340-1
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