Isolation and Characterization of Glycoproteins from 110 Day Old Ovine Fetal Diaphysial and Metaphysial Bone Fragments
Allswede, Michael P.
MetadataShow full item record
Glycoproteins, isolated from 110 day old fetal sheep compact and spongy bone samples, were fractionated and pooled according to order of elution from a molecular exclusion chromatography column. The major pool in the case of spongy and compact bone was investigated as to its homogeneity, amino acid composition, hexosamine composition, hexuronic acid composition, sialic acid composition, and neutral sugar composition. Techniques utilized to characterize the glycoproteins were molecular exclusion chromatography, colorimetric hexosamine assay, resorcinol assay, and gas chromatography. Glycoproteins isolated in this preparation were found to be of heterogeneous molecular weights and sizes. The glycoprotein fractions were, however, able to be pooled in 5-8 discreet pools based on similar protein, hexose, and hexuronic acid composition. The major pools of the compact and spongy bone samples were subjected to electrophoretic analysis to ascertain homogeneity, in those instances, 5-7 definable bands were seen to correspond between the two qlycoprotein preparations. The two major components (based on dye intensity) were also seen to correspond with one another. Further experimentation on the major oools of the two bone preparations was done to rouqhly characterize the glycoproteins in the pools and to aid in the further separation of these molecules. It was found that the protein in the molecules was composed of predominately serine, threonine, glycine, and alanine residues with a dearth of aromatic residues. The glycoproteins contained small amounts of sialic acid, hexosamine, and hexuronic acid. Larger amounts of hexose were found with galactose and mannose the predominate neutral sugars. Further separation is needed to accurately define the glycoproteins present in this preparation.