Fluorometric assay of neuraminidase with a sodium (4-methylumbelliferyl-α- d - N -acetylneuraminate) substrate

来自 dx.doi.org

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150

作者：

M. Potier a b c，L. Mameli a b c，M. Bélisle a b c，L. Dallaire a b c，S.B. Melanon a b c

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摘要：

This report describes the preparation of a sodium (4-methylumbelliferyl-α- d - N -acetylneuraminate) substrate and its use in a sensitive fluorometric assay of neuraminidase (EC 3.2.1.18) from Vibrio cholerae , cultured fibroblasts, and human leucocytes. V. cholerae neuraminidase showed maximum activity at pH 4.6 and an apparent K m of 1.5 m m and was activated by CaCl 2 and inhibited by ethylenediaminetetraacetate, NaCl, and N -acetylneuraminic acid. The inhibition by N -acetylneuraminic acid was competitive (K i = 6.1 m m ). Cultured fibroblast and leucocyte neuraminidases showed maximum activity between pH 4.2 and 4.4 and apparent K m values of 0.13 and 0.22 m m , respectively. Neuraminidase activity was considerably reduced in cultured fibroblasts of patients with mucolipidosis types I, II, and III.

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关键词：

Leukocytes Fibroblasts Humans Vibrio cholerae Sialic Acids Neuraminidase Spectrometry, Fluorescence

DOI：

10.1016/0003-2697(79)90362-2

被引量：

1583

年份：

1979