Effects of deglycosylation on the architecture of ovine submaxillary mucin glycoprotein.

The structural features of native and deglycosylated ovine submaxillary mucin (OSM) were determined by electron microscopy of platinum unidirectionally shadowed preparations and by ultracentrifugation. Thin filamentous molecules, of which 90% were 100-230 nm in length with estimated diameters of 1.0-1.4 nm, were observed with dilute samples of OSM in high ionic strength solvents (5-30 micrograms/ml in 0.8 M NaCl or NH4Ac). Ultracentrifugation studies indicated that these filamentous structures were monomers and/or dimers. At higher mucin concentrations or in lower ionic strength solvents, OSM molecules were oligomers that appeared as long rope-like strands. Removal of sialic acid residues by incubation with Clostridium perfringens neuraminidase yielded filamentous structures similar to those observed with OSM and some smaller less extended structures. Subsequent removal of the GalNAc residues of asialo-OSM with C. perfringens alpha-N-acetylgalactosaminidase resulted in a dramatic change in appearance, from an extended filament to a globular form. The frictional ratios of OSM and deglycosylated OSM were consistent with the marked structural differences of these molecules. Native OSM had a frictional ratio of 3.09, comparable to that of highly asymmetric tropomyosin (3.22); deglycosylated OSM had a frictional ratio of 1.11, comparable to that of globular ovalbumin (1.08).