Biochemical characterization of a sulfated phosphoglycoprotein antigen expressed on human small cell lung carcinoma.
A murine monoclonal antibody (mAb A23-16) was produced that recognizes a glycoprotein antigen preferentially expressed on the surface of human small cell lung carcinoma cells. This antibody is of IgG 1 isotype, has an association constant of 5 x 10(7) M-1, and reacts preferentially with human small cell lung carcinoma cell lines and fresh frozen sections in enzyme-linked immunosorbent assays and immunoperoxidase assays, respectively. The antigen recognized by A23-16 is a sulfated glycoprotein with phosphorylated threonine residues. The mature 90-kDa molecule has intrachain disulfide bonds and appears to be derived from a 76-kDa precursor, that is neither sulfated nor phosphorylated, but contains N-linked oligosaccharides. Conversion of the 76-kDa precursor to the mature form is accompanied by processing of these oligosaccharides from the high mannose to the complex type, although the increase in molecular mass from 76 to 90 kDa cannot be accounted for by this modification alone. MAb A23-16 reacts with its target antigen independent of the N-linked oligosaccharides, but requires intact intrachain disulfide bond(s) for reactivity. These studies on the molecular characterization of a monoclonal antibody-defined glycoprotein, preferentially expressed by small cell lung cancer, provide a basis for further structural and functional studies that may eventually lead to a delineation of its biological relevance for neoplastic transformation.
Koyama
Y
Y
Department of Immunology, Research Institute of Scripps Clinic, La Jolla, California 92037.
Yang
H M
HM
Wargalla
U
U
Reisfeld
R A
RA
Harper
J R
JR
D000328:Adult; D000911:Antibodies, Monoclonal; D018288:Carcinoma, Small Cell / Q000032:analysis*; D051152:Clusterin; D004797:Enzyme-Linked Immunosorbent Assay; D005333:Fetus / Q000032:analysis; D006023:Glycoproteins / Q000032:analysis*; D006596:Hexosaminidases / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D008175:Lung Neoplasms / Q000032:analysis*; D018832:Molecular Chaperones; D010766:Phosphorylation; D014018:Tissue Distribution; D014415:Tunicamycin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; C495242:CLU protein, human; D051152:Clusterin; D006023:Glycoproteins; D018832:Molecular Chaperones; D014415:Tunicamycin; D006596:Hexosaminidases