Detection of glycoproteins as tumor-associated Hanganutziu-Deicher antigen in human gastric cancer cell line, NUGC4.
NUGC4 cells derived from a human gastric cancer gave 6% Hanganutziu-Deicher (HD) antigen-positive cells by flow cytometric analysis using an affinity-purified chicken antibody to N-glycolyneuraminyl-lactosyl-ceramide (HD3 ganglioside). The cells showed no HD antigenic ganglioside by thin-layer chromatography enzyme-immunostaining; however, they were revealed to contain HD antigenic proteins with molecular masses of 150, 100, 90, 70, 65, 60, 47, and 40 kDa, by both immunoblotting after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoprecipitation of [35S]-methionine-labeled proteins, followed by SDS-PAGE and autoradiography. Neuraminidase treatment destroyed the antigenicity of all proteins, indicating that these molecules are glycoproteins and have N-glycolyneuraminic acid at the non-reducing terminal of carbohydrate chains as an HD antigenic epitope.
Fukui
Y
Y
Department of Pathology, Research Institute for Microbial Diseases, Osaka University, Japan.
Maru
M
M
Ohkawara
K
K
Miyake
T
T
Osada
Y
Y
Wang
D Q
DQ
Ito
T
T
Higashi
H
H
Naiki
M
M
Wakamiya
N
N
D015478:Antigens, Heterophile / Q000032:analysis*; D000951:Antigens, Neoplasm / Q000032:analysis*; D001345:Autoradiography; D002855:Chromatography, Thin Layer; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D005732:Gangliosides / Q000032:analysis; D006023:Glycoproteins / Q000032:analysis*; D006801:Humans; D015151:Immunoblotting; D007163:Immunosorbent Techniques; D008970:Molecular Weight; D009439:Neuraminidase / Q000494:pharmacology; D013274:Stomach Neoplasms / Q000276:immunology*; D014407:Tumor Cells, Cultured
D015478:Antigens, Heterophile; D000951:Antigens, Neoplasm; D005732:Gangliosides; D006023:Glycoproteins; D009439:Neuraminidase