Biotin binding to avidin. Oligosaccharide side chain not required for ligand association

Abstract

A commercially available, purified preparation of avidin was found to comprise two polypeptide bands (M(r) 18,000 and M(r) 15,500 respectively). Both bands bound biotin as assessed by biotin overlays of protein blots. The M(r) 15,500 polypeptide was found to differ from the M(r) 18,000 polypeptide only in its sugar content. When the commercial preparation was applied to a concanavalin A affinity column, the glycosylated forms were retarded as expected, and homotypic nonglycosylated avidin tetramers which failed to bind selectively to the column were collected in the effluent. The biotin-binding properties of the nonglycosylated avidin were equivalent to those obtained for the native (glycosylated) avidin molecule, indicating that the oligosaccharide moiety is not essential for the binding activity.