title: Identification of chitotriosidase isoforms in plasma of Gaucher disease patients by two dimensional gel electrophoresis.
abstract: 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.
authors.affiliation: Instituto Aragonés de Ciencias de la Salud, Edificio CEA, Avda. Gómez Laguna 25, 50009 Zaragoza.
authors.forename: Lucía
authors.initials: L
authors.lastname: Quintana
authors.forename: Alberto
authors.initials: A
authors.lastname: Monasterio
authors.forename: Kepa
authors.initials: K
authors.lastname: Escuredo
authors.forename: Jokin
authors.initials: J
authors.lastname: del Amo
authors.forename: Pilar
authors.initials: P
authors.lastname: Alfonso
authors.forename: Felix
authors.initials: F
authors.lastname: Elortza
authors.forename: Simon
authors.initials: S
authors.lastname: Santa Cruz
authors.forename: Laureano
authors.initials: L
authors.lastname: Simón
authors.forename: Antonio
authors.initials: A
authors.lastname: Martínez
authors.forename: Pilar
authors.initials: P
authors.lastname: Giraldo
authors.forename: Miguel
authors.initials: M
authors.lastname: Pocoví
authors.forename: José Luis
authors.initials: JL
authors.lastname: Castrillo
mesh_terms: D015153:Blotting, Western; D002384:Catalysis; D015180:Electrophoresis, Gel, Two-Dimensional; D005776:Gaucher Disease / Q000097:blood / Q000201:enzymology*; D006031:Glycosylation; D006596:Hexosaminidases / Q000032:analysis / Q000097:blood* / Q000737:chemistry; D006801:Humans; D007527:Isoenzymes / Q000032:analysis / Q000097:blood / Q000737:chemistry; D009439:Neuraminidase / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D007527:Isoenzymes; D006596:Hexosaminidases; C086400:chitotriosidase; D009439:Neuraminidase