title: Neuraminidase assay in cultured human fibroblasts: in situ versus in vitro procedures.
abstract: Further investigations have been carried out to characterize a published procedure of neuraminidase assay, in which the activity is measured directly on the cell culture layer. The pH optimum was 4.0. A Vmax value of 130 nmol/mg/h and a K(m) of 0.3 mmol/l were found. During incubation in the acid buffer, arylsulphatase activity was released into the medium, whereas neuraminidase activity remained attached to the cells. The in situ method allowed an unequivocal diagnosis of primary and secondary neuraminidase deficiencies. Because of its simplicity and reliability, the method appears useful as a routine method in clinical laboratories.
authors.affiliation: Kinderklinik, Universität Mainz, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Beck
authors.forename: E
authors.initials: E
authors.lastname: Scheuring
authors.forename: H U
authors.initials: HU
authors.lastname: Voelter
authors.forename: J
authors.initials: J
authors.lastname: Brandt
authors.forename: K
authors.initials: K
authors.lastname: Harzer
mesh_terms: D000328:Adult; D002478:Cells, Cultured; D002553:Cerebroside-Sulfatase / Q000032:analysis; D002648:Child; D002675:Child, Preschool; D005260:Female; D005347:Fibroblasts / Q000201:enzymology; D005456:Fluorescent Dyes; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D006923:Hymecromone / Q000031:analogs & derivatives; D007223:Infant; D007700:Kinetics; D008297:Male; D009439:Neuraminidase / Q000032:analysis / Q000172:deficiency / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes; D006923:Hymecromone; C021224:2'-(4-methylumbelliferyl)-alpha-D-N-acetylneuraminic acid; D002553:Cerebroside-Sulfatase; D009439:Neuraminidase