Evidence for sulfate modification of H-2Dd on N-linked carbohydrate(s): possible involvement in Ly-49A interaction.

Abstract

Murine class I molecules are ligands for Ly-49 molecules, a family of regulatory receptors expressed on murine NK cells. Since soluble sulfated mono- and polysaccharides interfere with the interaction of Ly-49A, a C-type lectin, and its class I ligand, Dd, it is possible that the oligosaccharides on class I molecules are sulfated and participate in Ly-49A binding. In this report, we show that H-2Dd expressed by activated T cells and various tumor cell lines is sulfated, as demonstrated by immunoprecipitation of Dd following Na235SO4 labeling. The 35SO4(-2) label on Dd expressed by a representative tumor cell, NZB1.1, is removed by peptide N-glycosidase F, but is resistant to endoglycosidase H treatment, indicating that the sulfate group is located on mature N-linked oligosaccharides. Two-dimensional SDS-PAGE analysis revealed that all major mature glycosylation variants of the Dd expressed by NZB1.1 are sulfated. Sodium chlorate, a potent inhibitor of ATP-sulfurylase, which prevents the formation of the sulfate donor, 3'-phosphoadenosine 5'-phosphosulfate, inhibited metabolic sulfation of Dd. NZB1.1 binds isolated Ly-49A immobilized on solid phase through an interaction by cell surface Dd, since cell adhesion was blocked by Abs directed against Dd or Ly-49A. Treatment of the Dd-expressing NZB1.1 tumor cells with sodium chlorate reduced their ability to bind immobilized Ly-49A, particularly when Ly-49A density was limiting. These results provide evidence for sulfation of H-2Dd oligosaccharide moieties, and suggest a role for this posttranslational modification in the interaction of Dd with Ly-49A.

• Authors: C S Chang, K P Kane
• Journal: Journal of immunology (Baltimore, Md. : 1950) (J Immunol) Vol. 160 Issue 9 Pg. 4367-74 (May 01 1998) ISSN: 0022-1767 [Print] United States
• PMID: 9574541 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antigens, Ly
• Carbohydrates
• H-2 Antigens
• Histocompatibility Antigen H-2D
• Lectins, C-Type
• Membrane Glycoproteins
• Receptors, Immunologic
• Receptors, NK Cell Lectin-Like
• Sulfates

Topics

• Animals
• Antigens, Ly
• Carbohydrates
• Electrophoresis, Gel, Two-Dimensional
• Female
• H-2 Antigens (immunology, metabolism)
• Histocompatibility Antigen H-2D
• Lectins, C-Type
• Lymphocyte Activation
• Membrane Glycoproteins (immunology, metabolism)
• Mice
• Mice, Inbred BALB C
• Receptors, Immunologic (immunology, metabolism)
• Receptors, NK Cell Lectin-Like
• Sulfates
• T-Lymphocytes (immunology, metabolism)
• Tumor Cells, Cultured