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

Proc Natl Acad Sci U S A. 2016 Apr 5;113(14):3791-6. doi: 10.1073/pnas.1514005113. Epub 2016 Mar 21.


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 cell-penetrating 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.

Keywords: GC activators; Gaucher's disease; LIMP-2; Parkinson's disease; β-glucocerebrosidase.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Motifs / physiology
  • Animals
  • Binding Sites
  • COS Cells
  • Cell Line
  • Chlorocebus aethiops
  • Crystallography, X-Ray
  • Glucosylceramidase / genetics
  • Glucosylceramidase / metabolism*
  • Humans
  • Lysosomal-Associated Membrane Protein 2 / genetics
  • Lysosomal-Associated Membrane Protein 2 / metabolism*
  • Lysosomes / metabolism*
  • Mice
  • Protein Binding
  • Protein Structure, Tertiary


  • Lysosomal-Associated Membrane Protein 2
  • Glucosylceramidase