Role of N-acetyl-D-galactosamine residue on B151K12-derived T cell-replacing factor (B151-TRF) molecule in B cell-receptor binding and -stimulating activity.
The role of sugar moiety on T cell-replacing factor molecule derived from a monoclonal T cell hybridoma B151K12 (B151-TRF) was analyzed with respect to the interaction with receptor on B cells. The induction of B cell differentiation into Ig-secreting cells by B151-TRF was specifically inhibited by addition of N-acetyl-D-galactosamine (GalNAc) to culture. Such inhibition appeared to be attributed to the interference of GalNAc in the interaction of TRF with its receptor, because absorption of TRF activity with B cells was notably inhibited by the presence of GalNAc. To substantiate this point further, we established binding assay of B151-TRF molecule to the receptor on B cells by using 125I-labeled TRF fraction enriched by reversed-phase high-performance liquid chromatography and gel filtration. The results revealed that the binding of 125I-TRF molecule to the B cells was almost completely blocked by GalNAc. Moreover, the existence of GalNAc residue(s) on B151-TRF molecule was evidenced by the facts that 1) the TRF activity was eluted from lectin gels with specificity for GalNAc as revealed by the functional assay, and 2) the 125I-TRF molecule specifically bound to such lectin gels. Thus, the GalNAc residue(s) on B151-TRF molecule plays an important role in binding of TRF molecule to the receptor and in the stimulation of B cells. The molecular properties of B cell-stimulatory B151-TRF and its mode of interaction with corresponding receptor on B cells were discussed in the context of B151-TRF as a glycosylated lymphokine molecule and B151-TRF receptor as a carbohydrate-binding protein (animal lectin).
D000042:Absorption; D000116:Acetylgalactosamine / Q000032:analysis / Q000494:pharmacology / Q000502:physiology*; D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D001667:Binding, Competitive; D005688:Galactosamine / Q000031:analogs & derivatives*; D006462:Hemolytic Plaque Technique; D006825:Hybridomas / Q000378:metabolism; D007166:Immunosuppressive Agents / Q000502:physiology; D015848:Interleukin-5; D007942:Leukemia, Experimental / Q000378:metabolism; D008213:Lymphocyte Activation; D008222:Lymphokines / Q000032:analysis / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D019158:N-Acetylneuraminic Acid; D011947:Receptors, Antigen, B-Cell / Q000378:metabolism* / Q000502:physiology; D012794:Sialic Acids / Q000032:analysis; D013601:T-Lymphocytes / Q000032:analysis / Q000378:metabolism*
D007166:Immunosuppressive Agents; D015848:Interleukin-5; D008222:Lymphokines; D011947:Receptors, Antigen, B-Cell; D012794:Sialic Acids; D005688:Galactosamine; D019158:N-Acetylneuraminic Acid; D000116:Acetylgalactosamine