Selective spin labeling of terminal galactose and N-acetylgalactosamine residues on the membrane surface of erythrocytes.

A method to selectively spin label galactose and N-acetylgalactosamine residues of erythrocyte membrane glycoconjugates is described. The method is based on the activation of the C-6 CH2OH group of these two sugars by galactose oxidase followed by reductive amination with 2,2,6,6-tetramethyl-4-aminopiperidine-1-oxyl in the presence of a mild reducing agent, NaBH3CN. The extent and distribution of the spin labeling suggest that the major sialoglycoprotein, PAS-1, incorporates the greatest amount of spin label while the glycolipids incorporate less than 10% of the spin label.