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Dissociation of glucocerebrosidase dimer in solution by its co-factor, saposin C

Journal Article · · Biochemical and Biophysical Research Communications
 [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1]
  1. National Inst. of Health (NIH), Bethesda, MD (United States)
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Mutations in the gene for the lysosomal enzyme glucocerebrosidase (GCase) cause Gaucher disease and are the most common risk factor for Parkinson disease (PD). Analytical ultracentrifugation of 8 μM GCase shows equilibrium between monomer and dimer forms. However, in the presence of its co-factor saposin C (Sap C), only monomer GCase is seen. Isothermal calorimetry confirms that Sap C associates with GCase in solution in a 1:1 complex (Kd = 2.1 ± 1.1 μM). Saturation cross-transfer NMR determined that the region of Sap C contacting GCase includes residues 63–66 and 74–76, which is distinct from the region known to enhance GCase activity. Because α-synuclein (α-syn), a protein closely associated with PD etiology, competes with Sap C for GCase binding, its interaction with GCase was also measured by ultracentrifugation and saturation cross-transfer. Unlike Sap C, binding of α-syn to GCase does not affect multimerization. However, adding α-syn reduces saturation cross-transfer from Sap C to GCase, confirming displacement. To explore where Sap C might disrupt multimeric GCase, GCase x-ray structures were analyzed using the program PISA, which predicted stable dimer and tetramer forms. In conclusion, for the most frequently predicted multimer interface, the GCase active sites are partially buried, suggesting that Sap C might disrupt the multimer by binding near the active site.
Research Organization:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Organization:
National Institutes of Health (NIH); USDOE
Grant/Contract Number:
W-31109-ENG-38
OSTI ID:
1225745
Alternate ID(s):
OSTI ID: 1233917
Journal Information:
Biochemical and Biophysical Research Communications, Journal Name: Biochemical and Biophysical Research Communications Journal Issue: 4 Vol. 457; ISSN 0006-291X
Publisher:
ElsevierCopyright Statement
Country of Publication:
United States
Language:
ENGLISH

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Cited By (4)

Did α-Synuclein and Glucocerebrosidase Coevolve? Implications for Parkinson’s Disease journal July 2015
A Review of Gaucher Disease Pathophysiology, Clinical Presentation and Treatments journal February 2017
Parkinson's disease: acid-glucocerebrosidase activity and alpha-synuclein clearance journal February 2016
Distinguishing the differences in β-glycosylceramidase folds, dynamics, and actions informs therapeutic uses journal October 2018

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
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Gaucher disease
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