Acta Pharm. 56 (2006) 95-104

Kinetics and activity of arylsulfatase A in leukocytes derived from patients with cerebral palsy

SVJETLANA KALANJ BOGNAR^1*,   BLAZENKA FORETIC², IVANA FURAC³,  ZELJKA VUKELIC²   and ZDRAVKO GRUBESIC⁴

svjetla@mef.hr

¹Croatian Institute for Brain Research, School of Medicine, University of Zagreb, Zagreb, Croatia
²Department of Chemistry and Biochemistry, School of Medicine, University of Zagreb, Zagreb, Croatia
³Department of Forensic Medicine and Criminology, School of Medicine, University of Zagreb, Zagreb, Croatia
⁴Special Children's Hospital for Neurodevelopmental Disorders "Goljak", Zagreb, Croatia

Activity and kinetics of arylsulfatase A (ASA, EC 3.1.6.8) were analyzed in leukocyte homogenates derived from patients suffering from cerebral palsy. Lower ASA activity was found in the patients' leukocytes than in controls, as determined by spectrophotometry using chromogenic substrate p-nitrocatechol sulfate (p-NCS). Kinetic parameters, Km and v^max, for leukocyte ASA were determined from the dependence of initial reaction velocities on the p-NCS concentrations. A slight difference in Km values was found for leukocyte enzyme in cerebral palsy (0.26 mmol L^-1) compared to the control (0.21 mmol L^-1), whereas max^-1 value for leukocyte ASA in disease reached only 58% of the control value. In addition, the presence of the most common mutations associated with ASA pseudodeficiency (N350S, 1524+95 A>G) and metachromatic leukodystrophy (P426L) was detected in all investigated patients. Changes in activity and kinetic parameters of leukocyte ASA in cerebral palsy are most probably related to the decrease of enzyme concentration; the detected mutations might at least partially contribute to the observed changes.

Keywords: arylsulfatase A, kinetic parameters, enzyme activity, mutations in ASA gene, cerebral palsy