Abstract
Asialoglycoprotein Ib and asialoglycocalicin have been isolated from the membranes and from the supernatant, respectively, after sonication of neuraminidase-treated platelets, by lectin affinity chromatography on peanut agglutinin. The isolated asialoglycoprotein Ib had an apparent molecular weight of 160,000 when not reduced and 150,000 when reduced, whereas the asialoglycocalicin had an apparent molecular weight of 150,000, both reduced and unreduced, on sodium dodecyl sulfate/polyacrylamide gel electrophoresis. Both preparations contained only trace amounts of impurities. The asialoglycoprotein Ib and asialoglycocalicin in both the unreduced and reduced states were separated by gel electrophoresis, radioiodinated in gel slices, and digested with trypsin, and the digests were analyzed by two-dimensional high-voltage electrophoresis and thin-layer chromatography followed by autoradiography. The tryptic peptide maps showed great similarities between glycocalicin and glycoprotein Ib, with the latter (both unreduced and reduced) containing additional peptides, supporting the idea that glycocalicin is derived from glycoprotein Ib. The unreduced glycoprotein Ib contained additional peptides compared to the reduced due to the disulfide-bond-linked beta component. There were also slight differences between unreduced and reduced glycocalicin, indicating that at least one intramolecular disulfide bond is present.