Identification of chitotriosidase isoforms in plasma of Gaucher disease patients by two dimensional gel electrophoresis.

Chitotriosidase protein (ChT) is the most important biochemical marker described for Gaucher disease (GD). ChT activity is increased several hundred-fold in plasma of GD patients and shows a strong positive correlation with the severity of the disease. However, a recessively inherited enzyme deficiency, with an incidence of about 6% in the Caucasian population, means that not all patients with GD can be monitored by measuring ChT activity. Applying two-dimensional gel electrophoresis (2-DE) technology this study describes the localization and identification of five ChT isoforms in 2-DE images obtained from plasma of GD patients. All these isoforms were unequivocally identified using MALDI-TOF mass spectrometry (MS) and validated by western blot analysis. The features of each ChT isoform separated by 2-DE in plasma from GD patients homozygous for the wild-type ChT allele, carriers of one defective allele and patients homozygous for the mutant allele are presented. We also show the correlation between each ChT isoform and the plasma ChT enzymatic activity of the GD patients sampled in this study.