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Analysis Note

Package sizes based on 4MU-NANA units

Package sizes based on the 4MU-NANA units

Application

Neuraminidase from Clostridium perfringens has been used in a study to assess purification via affinity chromatography. It has also been used in a study to investigate site-directed mutations of amino acids of the neuraminidase gene, nanH.

Biochem/physiol Actions

Neuraminidases are used to cleave terminal N-acetyl neuraminic acid (sialic acid) from a variety of glycoproteins. The enzyme from Clostridium perfringens cleaves terminal sialic acid residues which are α-2,3- α-2,6- or α-2,8-linked to Gal, GlcNac, GalNAc, AcNeu, GlcNeu, oligosaccharides, glycolipids or glycoproteins. The relative rate of cleavage decreases in the order: α-2-3 > α-2-6 . α-2-8. Neuraminidase from C. perfringens cleaves α-2-3 linked sialic acid residues most efficiently, compared to A. ureafaciens, (Sigma N3642) which preferentially cleaves α-2-6 linked residues.

Neuraminidase can block attachment of type 3 reovirus to cell membranes. This effect is related to the ability of neuraminidase to hydrolysis sialic acid residues within cell surface receptors.

The use of neuraminidase to remove sialic acid residues from glycoproteins on cell surfaces has been frequently reported. Generally, procedures have indicated using neuraminidase in PBS at 37°C for 30 minutes, followed by several washings with PBS. Treatment of tissue sections with neuraminidase at much lower concentrations require longer incubation: for 1-4 U/mL in 0.1 M acetate buffer pH 4.2-5, from 2 to 20 hours at 37 °C.

Neuraminidase cleavage of sialic acid groups has been used to study recognition by antibodies of glycoprotein structures. The use of neuraminidase in the estimation of N-acetylneuraminic acid was compared favorably to two other methods.

General description

Neuraminidase enzymes are glycoside hydrolase enzymes that catalyze hydrolysis of terminal sialic acid residues. The most well-known are the viral nearamidases, which promote influenza virus release.

Preparation Note

Purified by affinity chromatography from Type VIII (N 5631)

Unit Definition

One unit will hydrolyze 1.0 micromole of 2′-(4-Methylumbelliferyl)-alpha-D-N-actetylneuraminic acid per min at pH 5.0 at 37 °C.