Use of fluorescent substrates for characterization of Gaucher disease mutations

Idit Ron, Arie Dagan, Shimon Gatt, Metzada Pasmanik-Chor, Mia Horowitz*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


Gaucher disease results from impaired activity of the lysosomal enzyme β-glucocerebrosidase. More than 200 mutations within the glucocerebrosidase gene have been associated with this disease. In this study we tested the effect of several mutations (K157Q, D140H, E326K, D140H+E326K, V394L and R463C) on RNA stability, protein stability and activity toward four different fluorescent substrates (LR-12-GC, Bodipy-12-GC, LR-0-PAP-glucose and 4-MUG), using the vaccinia-derived expression system. The results indicated that the K157Q mutation leads to RNA instability, causing low protein levels and a concomitant reduction in β-glucocerebrosidase activity. All other tested mutations led to production of glucocerebrosidase RNA and protein with stabilities comparable to those of the normal counterpart. The D140H variant exhibited a high activity toward the tested substrates while the variant enzymes containing either the E326K or D140H and E326k mutations together expressed low β- glucocerebrosidase activity. The V394L variant exhibited low activity toward the tested substrates, while a higher activity was presented by the R463C containing glucocerebrosidase variant. Our results strongly indicated that the LR-12-GC substrate distinguishes between severities of different mutant glucocerebrosidase variants overexpressed in a heterologous system.

Original languageEnglish
Pages (from-to)57-65
Number of pages9
JournalBlood Cells, Molecules, and Diseases
Issue number1
StatePublished - Jul 2005


  • Fluorescent substrate
  • Gaucher disease
  • β-glucocerebrosidase


Dive into the research topics of 'Use of fluorescent substrates for characterization of Gaucher disease mutations'. Together they form a unique fingerprint.

Cite this