The adverse effect of neuraminidase on the analysis of cell surfaces by borohydride tritiation

• Published in: FEBS letters 1981-10, Vol.133 (1), p.165-168
• Main Authors: Thompson, S., Maddy, A.H.
• Format: Article
• Language: English
• Subjects: Adult; Borohydrides; carbohydrates; cell surface; Electrophoresis, Polyacrylamide Gel; Erythrocyte Membrane - analysis; Erythrocyte Membrane - drug effects; erythrocytes; Erythrocytes - analysis; Female; Galactose Oxidase; Glycoproteins - blood; Humans; Male; methodology; neuraminidase; Neuraminidase - pharmacology; Oxidation-Reduction; sodium borhydride; Tritium
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(81)80496-6

Many techniques have been developed to label surface components of cell membranes. One is the galactose oxidase/sodium borohydride procedure where terminal galactose or N-acetylgalactosamine residues of carbohydrate chains are first oxidised by galactose oxidase then tritiated by reduction with NaB super(3)H sub(4). In this method neuraminidase is almost invariably added along with the galactose oxidase as the removal of sialic acid increases the number of terminal galactose residues and hence the uptake of tritium on reduction. Using the erythrocyte surface as a model, the authors find that neuraminidase treatment has a deleterious effect on the mobility and resolution of labelled glycoproteins in sodium dodecyl sulphate (SDS)-polyacrylamide gel electrophoresis (PAGE). They suggest that this undesirable consequence of neuraminidase needs to be carefully considered when the enzyme is used to enhance galactose oxidase/borohydride tritiation of cell surfaces.