Zimmer G, Reuter G, Schauer R
Biochemisches Institut, Christian-Albrechts-Universität, Kiel, Federal Republic of Germany.
Eur J Biochem. 1992 Feb 15;204(1):209-15. doi: 10.1111/j.1432-1033.1992.tb16626.x.
An overlay and a solid-phase assay are presented which allow the specific detection of 9-O-acetylated sialic acids on sialoglycoconjugates immobilized on microtiter plates, nitrocellulose or separated on thin-layer chromatograms. The assay takes advantage of two different biological properties of influenza C virus, its high-affinity binding to 9-O-acetylated sialic acids and its sialate 9-O-acetylesterase that is used for detection of bound virus with fluorogenic or chromogenic substrates. Though simple and rapid, the assay is highly sensitive with a detection limit of 65 fmol 9-O-acetylated sialic acid in 9-O-acetylated ganglioside GD1a. Influenza C virus is able to bind to a wide spectrum of sialoglycoconjugates like mucins, serum glycoproteins or gangliosides containing naturally or synthetically O-acetylated sialic acids. 9-O-Acetyl-N-glycoloylneuraminic acid can also function as a high-affinity receptor determinant for influenza C virus. While the acetyl ester at the 9 position is essential for virus binding in all cases, a 4-O-acetyl group is not recognized. In addition to alpha(2.3) or alpha(2.6) bonds, 9-O-acetyl-N-acetylneuraminic acid in alpha(2.8) linkage to N-acetylneuraminic acid is also functionally active.
本文介绍了一种覆盖法和一种固相分析法，可用于特异性检测固定在微量滴定板、硝酸纤维素膜上的唾液酸糖缀合物上的9-O-乙酰化唾液酸，或在薄层色谱上分离的唾液酸糖缀合物上的9-O-乙酰化唾液酸。该分析利用了丙型流感病毒的两种不同生物学特性，即其与9-O-乙酰化唾液酸的高亲和力结合以及其唾液酸9-O-乙酰酯酶，该酶用于用荧光或显色底物检测结合的病毒。尽管该分析简单快速，但灵敏度很高，在9-O-乙酰化神经节苷脂GD1a中检测限为65 fmol 9-O-乙酰化唾液酸。丙型流感病毒能够结合多种唾液酸糖缀合物, 如粘蛋白、血清糖蛋白或含有天然或合成O-乙酰化唾液酸的神经节苷脂。9-O-乙酰基-N-糖基神经氨酸也可作为丙型流感病毒的高亲和力受体决定簇。虽然在所有情况下9位的乙酰酯对于病毒结合至关重要，但4-O-乙酰基不被识别。除了α(2,3)或α(2,6)键外，以α(2,8)连接到N-乙酰神经氨酸的9-O-乙酰基-N-乙酰神经氨酸也具有功能活性。
