Isolation and characterization of leukosialin, a major sialoglycoprotein on human leukocytes.
A major sialoglycoprotein (previously called gp105) on the human erythroleukemic cell line K562 was purified, and specific antibodies were raised in a rabbit. A number of different hematopoietic cell lines belonging to erythroid, myeloid, T-lymphoid, and B-lymphoid cell lineages were found to possess glycoproteins that were immunoprecipitated by these antibodies. However, the apparent molecular weights differed between cell lines, ranging from 113,000 to 150,000. In almost all cases, the immune precipitated molecule corresponded to the major sialoglycoprotein of the respective cell. Pulse-chase experiments showed that all cells produced an early precursor form of the molecule of 54 kDa, which was susceptible to endo-beta-N-acetylglucosaminidase H to give an apoprotein of 52 kDa. Neuraminidase treatment of the mature forms resulted in a characteristic decrease of the mobility in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (apparent molecular weights from 150,000 to 183,000). Amino acid analysis of the glycoprotein isolated from HL-60 cells showed a high content of serine, threonine, and proline, and the carbohydrate composition was compatible with the presence of a large number (approximately 90) of O-linked carbohydrate chains. The name leukosialin is proposed for this sialoglycoprotein, which seems to be widely distributed, but differently glycosylated, on leukocytes with diverse functions. In the following paper (Carlsson, S.R., Sasaki, H., and Fukuda, M. (1986) J. Biol. Chem. 261, 12787-12795), we demonstrate that the structures of O-linked oligosaccharides vary significantly depending on the cells from which leukosialin was isolated.
D000818:Animals; D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D002460:Cell Line; D005944:Glucosamine / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000032:analysis; D006596:Hexosaminidases / Q000378:metabolism; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000032:analysis; D015470:Leukemia, Myeloid, Acute / Q000378:metabolism; D007962:Leukocytes / Q000032:analysis*; D051917:Leukosialin; D008715:Methionine / Q000378:metabolism; D008970:Molecular Weight; D011402:Pronase / Q000378:metabolism; D011817:Rabbits; D012795:Sialoglycoproteins / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D051917:Leukosialin; C465014:SPN protein, human; D012795:Sialoglycoproteins; D008715:Methionine; D006596:Hexosaminidases; D011402:Pronase; D005944:Glucosamine