Identification and structural characterization of Lyt-1 glycoproteins from tunicate hemocytes and mouse thymocytes.
1. A panel of monoclonal antibodies specific to murine Lyt-1 allotypic and framework determinants was used to investigate the possible occurrence of a Lyt-1 homolog in tunicate (protochoradte) hemocytes. 2. In immunoprecipitation experiments, antigenic activities were associated with a major 67 kDa component on tunicate hemocytes and C57Bl/6 mouse thymocytes. 3. Tunicate and mouse Lyt-1 molecules were compared, in terms of glycosylation, by their sensitivity to glycosidases and analyses on one- and two-dimensional gel electrophoresis. 4. Each of the two molecules appeared to bear two N-linked oligosaccharides, one high-mannose and one complex-type glycan. 5. Both molecules revealed charge microheterogeneity with differences in sialic acid content accounting for the charge difference between each other. 6. However, the difference in the glycans did not account for the microheterogeneity within each molecule, suggesting that other post-translational modifications might be responsible. 7. At the polypeptide level, comparisons of chymotryptic and endoproteinase-Arg-C peptide maps, as well as CNBr-cleavage products, suggested that tunicate and mouse Lyt-1 molecules are structurally similar and that each may contain at least one intra-chain disulfide bridge. 8. The significance of these findings is discussed in terms of the possible biological role of Lyt-1 glycoproteins at different levels of evolution.
Negm
H I
HI
Department of Anatomy, School of Medicine, University of California, Los Angeles 90024.
Mansour
M H
MH
Cooper
E L
EL
D000818:Animals; D000911:Antibodies, Monoclonal; D005075:Biological Evolution; D002241:Carbohydrates / Q000737:chemistry; D003488:Cyanogen Bromide; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D007518:Isoantibodies / Q000097:blood / Q000737:chemistry*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010449:Peptide Mapping; D011233:Precipitin Tests; D013950:Thymus Gland / Q000737:chemistry* / Q000166:cytology; D014561:Urochordata
D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D007518:Isoantibodies; C029805:Lyt antibodies; D003488:Cyanogen Bromide