Characterization of the complex formed by β-glucocerebrosidase and the lysosomal integral membrane protein type-2

Friederike Zunke, Lisa Andresen, Sophia Wesseler, Johann Groth, Philipp Arnold, Michelle Rothaug, Joseph R. Mazzulli, Dimitri Krainc, Judith Blanz, Paul Saftig, Michael Schwake*

*Corresponding author for this work

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

The lysosomal integral membrane protein type-2 (LIMP-2) plays a pivotal role in the delivery of β-glucocerebrosidase (GC) to lysosomes. Mutations in GC result in Gaucher's disease (GD) and are the major genetic risk factor for the development of Parkinson's disease (PD). Variants in the LIMP-2 gene cause action myoclonus-renal failure syndrome and also have been linked to PD. Given the importance of GC and LIMP-2 in disease pathogenesis, we studied their interaction sites in more detail. Our previous data demonstrated that the crystal structure of LIMP-2 displays a hydrophobic three-helix bundle composed of helices 4, 5, and 7, of which helix 5 and 7 are important for ligand binding. Here, we identified a similar helical motif in GC through surface potential analysis. Coimmunoprecipitation and immunofluorescence studies revealed a triple-helical interface region within GC as critical for LIMP-2 binding and lysosomal transport. Based on these findings, we generated a LIMP-2 helix 5-derived peptide that precipitated and activated recombinant wild-type and GD-associated N370S mutant GC in vitro. The helix 5 peptide fused to a cellpenetrating peptide also activated endogenous lysosomal GC and reduced α-synuclein levels, suggesting that LIMP-2-derived peptides can be used to activate endogenous as well as recombinant wild-type or mutant GC efficiently. Our data also provide a structural model of the LIMP-2/GC complex that will facilitate the development of GC chaperones and activators as potential therapeutics for GD, PD, and related synucleinopathies.

Original languageEnglish (US)
Pages (from-to)3791-3796
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume113
Issue number14
DOIs
StatePublished - Apr 5 2016

Fingerprint

Lysosome-Associated Membrane Glycoproteins
Glucosylceramidase
Gaucher Disease
Parkinson Disease
Peptides
Synucleins
Progressive Myoclonic Epilepsy
Structural Models
Lysosomes
Protein Binding
Fluorescent Antibody Technique

Keywords

  • GC activators
  • Gaucher's disease
  • LIMP-2
  • Parkinson's disease
  • β-glucocerebrosidase

ASJC Scopus subject areas

  • General

Cite this

Zunke, Friederike ; Andresen, Lisa ; Wesseler, Sophia ; Groth, Johann ; Arnold, Philipp ; Rothaug, Michelle ; Mazzulli, Joseph R. ; Krainc, Dimitri ; Blanz, Judith ; Saftig, Paul ; Schwake, Michael. / Characterization of the complex formed by β-glucocerebrosidase and the lysosomal integral membrane protein type-2. In: Proceedings of the National Academy of Sciences of the United States of America. 2016 ; Vol. 113, No. 14. pp. 3791-3796.
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Characterization of the complex formed by β-glucocerebrosidase and the lysosomal integral membrane protein type-2. / Zunke, Friederike; Andresen, Lisa; Wesseler, Sophia; Groth, Johann; Arnold, Philipp; Rothaug, Michelle; Mazzulli, Joseph R.; Krainc, Dimitri; Blanz, Judith; Saftig, Paul; Schwake, Michael.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 113, No. 14, 05.04.2016, p. 3791-3796.

Research output: Contribution to journalArticle

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T1 - Characterization of the complex formed by β-glucocerebrosidase and the lysosomal integral membrane protein type-2

AU - Zunke, Friederike

AU - Andresen, Lisa

AU - Wesseler, Sophia

AU - Groth, Johann

AU - Arnold, Philipp

AU - Rothaug, Michelle

AU - Mazzulli, Joseph R.

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AU - Blanz, Judith

AU - Saftig, Paul

AU - Schwake, Michael

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AB - The lysosomal integral membrane protein type-2 (LIMP-2) plays a pivotal role in the delivery of β-glucocerebrosidase (GC) to lysosomes. Mutations in GC result in Gaucher's disease (GD) and are the major genetic risk factor for the development of Parkinson's disease (PD). Variants in the LIMP-2 gene cause action myoclonus-renal failure syndrome and also have been linked to PD. Given the importance of GC and LIMP-2 in disease pathogenesis, we studied their interaction sites in more detail. Our previous data demonstrated that the crystal structure of LIMP-2 displays a hydrophobic three-helix bundle composed of helices 4, 5, and 7, of which helix 5 and 7 are important for ligand binding. Here, we identified a similar helical motif in GC through surface potential analysis. Coimmunoprecipitation and immunofluorescence studies revealed a triple-helical interface region within GC as critical for LIMP-2 binding and lysosomal transport. Based on these findings, we generated a LIMP-2 helix 5-derived peptide that precipitated and activated recombinant wild-type and GD-associated N370S mutant GC in vitro. The helix 5 peptide fused to a cellpenetrating peptide also activated endogenous lysosomal GC and reduced α-synuclein levels, suggesting that LIMP-2-derived peptides can be used to activate endogenous as well as recombinant wild-type or mutant GC efficiently. Our data also provide a structural model of the LIMP-2/GC complex that will facilitate the development of GC chaperones and activators as potential therapeutics for GD, PD, and related synucleinopathies.

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