Characterization of a unique glycoprotein antigen expressed on the surface of human neuroblastoma cells

K. Mujoo, R. C. Spiro, R. A. Reisfeld

Research output: Contribution to journalArticle

34 Scopus citations

Abstract

In order to develop a molecular probe to delineate chemical and biological characteristics of human neuroblastoma cells, a murine monoclonal antibody (Mab 5G3) was produced that is directed to a glycoprotein, preferentially expressed on the surface of such cells. This antibody is of IgG2a isotype, has an association constant of 8 x 109 M-1, and reacts preferentially with human neuroblastoma cell lines and fresh frozen tissue sections in enzyme-linked immunosorbent assay and immunoperoxidase assays, respectively. Minimal reactivity is observed with a variety of lymphoblastoid cell lines and normal fetal and adult tissues. Mab 5G3 specifically recognizes a neuroblastoma target glycoprotein antigen of 215 kDa that is derived form a 200-kDa precursor, as evident from pulse-chase biosynthetic studies. Treatment with tunicamycin revealed that both molecules contain N-asparagine-linked oligosaccharides; however, only the 215-kDa species is resistant to treatment with endo-β-acetylglucosaminidase H and sensitive to neuroaminidase, indicating that it contains trimmed and terminally sialylated oligosaccharides of the 'complex' type. In contrast, the 200-kDa precursor is sensitive to endo-β-N-acetylglucosaminidase H and resistant to neuraminidase treatment indicating that it contains high-mannose nonprocessed oligosaccharides. The 215-kDa molecule is sulfated, phosphorylated at serine residues, and expressed on the cell surface. A molecule of 200 kDa is detected by Mab 5G3 in spent culture medium of human neuroblastoma cells which is neither sulfated nor phosphorylated.

Original languageEnglish (US)
Pages (from-to)10299-10305
Number of pages7
JournalJournal of Biological Chemistry
Volume261
Issue number22
StatePublished - 1986

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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