FLUOROMETRIC ASSAY OF NEURAMINIDASE WITH A SODIUM (4-METHYLUMBELLIFERYL-ALPHA-D-N-ACETYLNEURAMINATE) SUBSTRATE
FLUOROMETRIC ASSAY OF NEURAMINIDASE WITH A SODIUM (4-METHYLUMBELLIFERYL-ALPHA-D-N-ACETYLNEURAMINATE) SUBSTRATE
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DOI：
10.1016/0003-2697(79)90362-2
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发表时间：
1979-01-01
影响因子：
2.9
通讯作者：
MELANCON, SB
中科院分区：
文献类型：
--
作者：
POTIER, M;MAMELI, L;MELANCON, SB
（4-甲基伞形酮基-α-）钠的制备D-N-乙酰神经氨酸）底物及其在来自霍乱弧菌、培养的成纤维细胞和人白细胞的神经氨酸酶（EC3.2.1.18）的灵敏荧光测定中的用途。霍乱弧菌神经氨酸酶在pH 4.6和1.5 mM的表观Km下显示出最大活性，并且被CaCl 2激活，并被EDTA、NaCl和N-乙酰神经氨酸抑制。N-乙酰神经氨酸的抑制是竞争性的（Ki [抑制常数] = 6.1 mM）。培养的成纤维细胞和白细胞神经氨酸酶显示出最大活性之间的pH 4.2-4.4和表观Km值分别为0.13和0.22 mM。神经氨酸酶活性显着降低，在培养的成纤维细胞的粘脂沉积症I型，II型和III型的患者。
The preparation of a sodium (4-methylumbelliferyl-.alpha.-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 leukocytes is described. V. cholerae neuraminidase showed maximum activity at pH 4.6 and an apparent Km of 1.5 mM, and was activated by CaCl2 and inhibited by EDTA, NaCl, and N-acetylneuraminic acid. The inhibition by N-acetylneuraminic acid was competitive (Ki [inhibition constant] = 6.1 mM). Cultured fibroblast and leukocyte neuraminidases showed maximum activity between pH 4.2-4.4 and apparent Km values of 0.13 and 0.22 mM, respectively. Neuraminidase activity was considerably reduced in cultured fibroblasts of patients with mucolipidosis types I, II and III.