title: A rapid fluorogenic palmitoyl-protein thioesterase assay: pre- and postnatal diagnosis of INCL.
abstract: A deficiency of palmitoyl-protein thioesterase (PPT) was recently shown to be the primary defect in infantile neuronal ceroid lipofuscinosis (INCL). The available enzyme assays are complicated and impractical for diagnostic use. We have recently developed a new, fluorometric assay for PPT based on the sensitive fluorochrome 4-methylumbelliferone, requiring an overnight incubation to measure PPT. Now we have synthesized an analogue of this substrate which allows PPT determinations in 1 h. This improved PPT assay is simple, sensitive, and robust and will facilitate the definition of the full clinical spectrum associated with a deficiency of PPT. PPT activity was readily detectable in fibroblasts, leukocytes, amniotic fluid cells, chorionic villi, plasma, and cerebrospinal fluid from controls. PPT activity was profoundly deficient in these tissues and fluids from INCL patients. Similarly, a deficiency of PPT activity was demonstrated in patients with the variant juvenile NCL with GROD. These results show the feasibility of rapid pre- and postnatal diagnosis of INCL and its variants.
authors: Department of Clinical Genetics, Erasmus University, 3000 DR, Rotterdam, The Netherlands. vandiggelen@kgen.fgg.eur.nl
authors: O P
authors: OP
authors: van Diggelen
authors: J L
authors: JL
authors: Keulemans
authors: B
authors: B
authors: Winchester
authors: I L
authors: IL
authors: Hofman
authors: S L
authors: SL
authors: Vanhanen
authors: P
authors: P
authors: Santavuori
authors: Y V
authors: YV
authors: Voznyi
mesh_terms: D004796:Clinical Enzyme Tests; D004305:Dose-Response Relationship, Drug; D005470:Fluorometry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009472:Neuronal Ceroid-Lipofuscinoses / Q000175:diagnosis*; D011296:Prenatal Diagnosis / Q000379:methods*; D013869:Thiolester Hydrolases / Q000032:analysis* / Q000097:blood / Q000134:cerebrospinal fluid; D013997:Time Factors
chemical_list: D013869:Thiolester Hydrolases; C095299:palmitoyl-protein thioesterase