Characterization of gp39, a B-lymphocyte associated differentiation antigen which is also present on granulocytes and macrophages.
The biosynthesis and biochemical characteristics of the 39,000 cell surface glycoprotein detected by Mab 41H.16 were investigated. Experiments utilizing tunicamycin, endoglycosidase H, endoglycosidase F and N-glycosidase F indicate that the mature molecule expressed at the cell surface is composed largely of N-linked oligosaccharides of both the complex and high mannose types. When synthesized in the presence of tunicamycin, the molecule appeared on the cell surface with a Mr of 32,000. Digestion with both endoglycosidase H and endoglycosidase F yielded a single band of Mr 37,000. Parallel experiments with N-glycosidase F revealed species of approx. 35,000 and 32,000. Synthesis in the presence of monensin yielded a 37,500 product. [3H]Glucosamine and [3H]mannose were incorporated into the molecule but no evidence for fucose incorporation could be found. Microheterogeneity of gp39 with respect to Mr and oligosaccharide structure was demonstrated by biosynthetic labelling and lectin chromatography. Biosynthetic pulse-chase labelling showed that the de novo synthesis of the 39,000 molecule occurs without detectable precursor formation. Results of temperature-dependent phase separation experiments were consistent with gp39 being an integral membrane protein. Two-dimensional electrophoresis showed heterogeneity of the isoelectric points associated with the N-linked oligosaccharides. Galactose oxidase/NaB[3H]4 labelling showed that a terminal sialic acid protects a galactose residue. All results are consistent with the conclusion that the gp39 molecule is an integral membrane glycoprotein composed of heterogeneous N-linked oligosaccharides of both the complex and high mannose types.
Lauzon
G J
GJ
Department of Pediatrics, University of Calgary, Alberta, Canada.
Antoun
G R
GR
Longenecker
B M
BM
Zipf
T F
TF
D000944:Antigens, Differentiation, B-Lymphocyte / Q000096:biosynthesis / Q000302:isolation & purification*; D001402:B-Lymphocytes / Q000276:immunology*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D006098:Granulocytes / Q000276:immunology*; D008264:Macrophages / Q000276:immunology*; D008985:Monensin / Q000494:pharmacology; D014415:Tunicamycin / Q000494:pharmacology
D000944:Antigens, Differentiation, B-Lymphocyte; D014415:Tunicamycin; D008985:Monensin