Molecular cloning of the 31 kDa cytosolic phospholipase A2, as an antigen recognized by the lung cancer-specific human monoclonal antibody, AE6F4.
The human monoclonal antibody AE6F4 specifically reacts with human lung cancer tissues but does not with normal tissues. This monoclonal antibody recognizes a cytosolic 31 kDa antigen in the cancer cells. In a previous study, we elucidated that the 31 kDa antigen belonged to a family of proteins collectively designated as 14-3-3 proteins, which were known as protein kinase-dependent activators of tyrosine/trytophan hydroxylases, or protein kinase C inhibitor proteins. Here we report molecular cloning of the 31 kDa antigen from the human lung adenocarcinoma cell line, A549. Sequencing analysis indicates that the cloned cDNA is identical to that of previously reported human placental cytosolic phospholipase A2 (cPLA2), which is also a member of the 14-3-3 protein family. Western analysis demonstrated that a 31 kDa recombinant cPLA2 expressed in monkey COS cells was recognized by the AE6F4 monoclonal antibody. Binding of the monoclonal antibody to the recombinant cPLA2 was abolished when treated with sodium periodate, suggesting that not only are carbohydrate chains associated with the cPLA2, but they also play a crucial role in antigen recognition by the monoclonal antibody.
Kawamoto
S
S
Graduate School of Genetic Resources Technology, Kyushu University, Fukuoka, Japan.
Shoji
M
M
Setoguchi
Y
Y
Kato
M
M
Hashizume
S
S
Ichikawa
A
A
Osada
K
K
Katakura
Y
Y
Tachibana
H
H
Murakami
H
H
D048948:14-3-3 Proteins; D000230:Adenocarcinoma / Q000276:immunology* / Q000473:pathology; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000912:Antibodies, Neoplasm / Q000276:immunology*; D000918:Antibody Specificity; D000951:Antigens, Neoplasm / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular; D003600:Cytosol / Q000201:enzymology; D018076:DNA, Complementary / Q000032:analysis; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D008175:Lung Neoplasms / Q000276:immunology* / Q000473:pathology; D008969:Molecular Sequence Data; D010741:Phospholipases A / Q000235:genetics* / Q000276:immunology; D054467:Phospholipases A2; D011506:Proteins / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D014407:Tumor Cells, Cultured; D014446:Tyrosine 3-Monooxygenase
D048948:14-3-3 Proteins; D000911:Antibodies, Monoclonal; D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm; D015295:Antigens, Tumor-Associated, Carbohydrate; D018076:DNA, Complementary; D011506:Proteins; D012333:RNA, Messenger; D014446:Tyrosine 3-Monooxygenase; D010741:Phospholipases A; D054467:Phospholipases A2