Human Golgi β‐galactoside α‐2,6‐sialyltransferase generates a group of sialylated B lymphocyte differentiation antigens

The role of the human β‐galactoside α‐2,6‐sialyltransferase (hu α‐2,6‐ST) in the generation of B cell surface antigens was investigated by selecting subclones of COS cells (monkey kidney epithelial cells) constitutively expressing a transfected cDNA which encodes the hu α‐2,6‐ST (COS α‐2,6‐ST cells). Expression of hu α‐2,6‐ST in COS cells was sufficient to generate sialylated cell surface epitopes on different glycosylated antigens recognized by monoclonal antibodies to CDw75, CD76, and the unclustered monoclonal antibodies HB‐4 and EBU‐65. These epitopes were sensitive to sialidase treatment and are likely to contain terminal α‐2,6‐linked sialic acid residues. A novel antiserum raised against bacterially expessed hu α‐2,6‐ST fusion protein was used to localize the sialyltransferase in two cell lines with high expression of either endogenous (B cell line JOK‐1) or recombinant (COS α‐2,6‐ST cells) hu α‐2,6‐ST. In both cell lines, the enzyme was detected only intracellularly in the juxtanuclear region and not on the cell surface. In contrast, CDw75, formerly proposed to be identical with an α‐2,6‐ectosialyltransferase, was strongly expressed on the cell surface. The different expression patterns show that neither the CDw75 antigen nor any of the other sialylated antigens analyzed is identical with the hu α‐2,6‐ST. Furthermore, the presence of a surface‐expressed α‐2,6‐ST appears unlikely in these cell lines. We propose that CDw75, CD76, HB‐4, and EBU‐65 represent a unique group of B cell differentiation antigens the production of which requires the enzymatic activity of α‐2,6‐ST.

[1]  S. Wong,et al.  The 17-residue transmembrane domain of beta-galactoside alpha 2,6- sialyltransferase is sufficient for Golgi retention , 1992, The Journal of cell biology.

[2]  S. Munro,et al.  The B lymphocyte surface antigen CD75 is not an α-2,6-sialyltransferase but is a carbohydrate antigen, the production of which requires the enzyme , 1992, Cell.

[3]  G. Freeman,et al.  The HB-6, CDw75, and CD76 differentiation antigens are unique cell- surface carbohydrate determinants generated by the beta-galactoside alpha 2,6-sialyltransferase , 1992, The Journal of cell biology.

[4]  S. Munro Sequences within and adjacent to the transmembrane segment of alpha-2,6-sialyltransferase specify Golgi retention. , 1991, The EMBO journal.

[5]  P. Möller,et al.  Rapid intracellular pathway gives rise to cell surface expression of the MHC class II-associated invariant chain (CD74). , 1991, Journal of immunology.

[6]  K. Guy,et al.  Expression of the CDw75 (beta-galactoside alpha 2,6-sialyltransferase) antigen on normal blood cells and in B-cell chronic lymphocytic leukaemia. , 1991, Immunology.

[7]  D. Sgroi,et al.  The B lymphocyte adhesion molecule CD22 interacts with leukocyte common antigen CD45RO on T cells and α2–6 sialyltransferase, CD75, on B cells , 1991, Cell.

[8]  S. Evans,et al.  Evidence for a molecular distinction between Golgi and cell surface forms of beta 1,4-galactosyltransferase. , 1991, The Journal of biological chemistry.

[9]  B. Dörken,et al.  CD22 antigen: biosynthesis, glycosylation and surface expression of a B lymphocyte protein involved in B cell activation and adhesion. , 1991, International immunology.

[10]  E. Smeland,et al.  The B cell antigen CD75 is a cell surface sialytransferase , 1990, The Journal of experimental medicine.

[11]  B. Dörken,et al.  Monoclonal antibodies against the human lymphocyte differentiation antigen CD 76 bind to gangliosides , 1990, FEBS letters.

[12]  J. Paulson,et al.  Conversion of a Golgi apparatus sialyltransferase to a secretory protein by replacement of the NH2-terminal signal anchor with a signal peptide. , 1989, The Journal of biological chemistry.

[13]  J. Paulson,et al.  Glycosyltransferases. Structure, localization, and control of cell type-specific glycosylation. , 1989, The Journal of biological chemistry.

[14]  R. Döffinger,et al.  Electrotransfection of human lymphoid and myeloid cell lines. , 1988, Nucleic acids research.

[15]  J. Paulson,et al.  Post-Golgi apparatus localization and regional expression of rat intestinal sialyltransferase detected by immunoelectron microscopy with polypeptide epitope-purified antibody. , 1988, The Journal of biological chemistry.

[16]  R. Cummings,et al.  The immobilized leukoagglutinin from the seeds of Maackia amurensis binds with high affinity to complex-type Asn-linked oligosaccharides containing terminal sialic acid-linked alpha-2,3 to penultimate galactose residues. , 1988, The Journal of biological chemistry.

[17]  M. Thurnher,et al.  Galactosyltransferase and sialyltransferase are located in different subcellular compartments in HeLa cells. , 1987, Experimental cell research.

[18]  G. Hart,et al.  Cell surface sialic acid influences tumor cell recognition in the mixed lymphocyte reaction. , 1987, Journal of immunology.

[19]  J. Gerdes,et al.  Novel type of proliferating lymphoplasmacytoid cell with a characteristic spotted immunofluorescence pattern. , 1987, The Journal of clinical investigation.

[20]  I. Goldstein,et al.  The elderberry (Sambucus nigra L.) bark lectin recognizes the Neu5Ac(alpha 2-6)Gal/GalNAc sequence. , 1987, The Journal of biological chemistry.

[21]  H. Nariuchi,et al.  The regulatory role of sialic acids in the response of class II reactive T cell hybridomas to allogeneic B cells. , 1986, Journal of immunology.

[22]  V. Diehl,et al.  Unique antigen of cultured Hodgkin's cells. A putative sialyltransferase. , 1986, The Journal of clinical investigation.

[23]  Roland Schauer,et al.  Sialic acids and their role as biological masks , 1985 .

[24]  M. Cooper,et al.  Development and distribution of a human B cell subpopulation identified by the HB-4 monoclonal antibody. , 1985, Journal of immunology.

[25]  R. Massarelli,et al.  Ectoglycosyltransferase activities at the surface of cultured neurons. , 1983, Biochemical and biophysical research communications.

[26]  B. Bauvois,et al.  Membrane glycoprotein IIb is the major endogenous acceptor for human platelet ectosialyltransferase , 1981, FEBS letters.

[27]  Y. Gluzman SV40-transformed simian cells support the replication of early SV40 mutants , 1981, Cell.

[28]  S. Roseman,et al.  EVIDENCE FOR CELL-SURFACE GLYCOSYLTRANSFERASES , 1971, The Journal of cell biology.