Isolation and characterization of a mannose/N-acetylglucosamine/fucose-binding protein from rat liver

A rat liver mannan-binding protein was isolated by affinity chromatography on invertase--Sepharose by a modification of the method of Kawasaki, Etoh & Yamashina [(1978) Biochem. Biophys. Res. Commun. 81, 1018-1024] and by a new method involving chromatography on mannose-Sepharose. The binding protein appears as a single band on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis with an apparent mol.wt. of approx. 30000. Binding of 125I-labelled mannan is saturable and inhibited by mannose, N-acetylglucosamine, or L-fucose but not by galactose or mannose 6-phosphate. Neoglycoproteins containing mannose, N-acetylglucosamine, or L-fucose, but not galactose, are inhibitory. The neoglycoproteins are 10000-fold more effective (based on moles of sugar) than are free monosaccharides as inhibitors. 125I-labelled mannan binding to the binding protein is calcium-dependent.