Identification and intracellular location of MAGE-3 gene product.
The human MAGE-3 gene encodes a melanoma antigenic epitope recognized by specific cytotoxic T lymphocytes, but its gene product has not been identified thus far. We produced a recombinant MAGE-3 gene product by expression cloning of the entire reading frame in the context of a fusion protein characterized by a 10-histidine tail, allowing purification by metal chelation on a nickel Sepharose column. The semipurified product was used to generate MAGE-3-specific monoclonal antibodies. One reagent could identify by immunoblotting the native MAGE-3 gene product as a M(r) 48,000 protein in lysates of cell lines showing evidence of MAGE-3 gene expression. No apparent cross-reactivity with recombinant or native MAGE-1 gene product was observed. Immunohistochemistry shows that, closely resembling the MAGE-1 gene product, MAGE-3 is a cytoplasmic protein.
Kocher
T
T
Department of Surgery, University of Basel, Switzerland.
Schultz-Thater
E
E
Gudat
F
F
Schaefer
C
C
Casorati
G
G
Juretic
A
A
Willimann
T
T
Harder
F
F
Heberer
M
M
Spagnoli
G C
GC
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000951:Antigens, Neoplasm / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular; D015870:Gene Expression; D007424:Intracellular Fluid / Q000737:chemistry / Q000276:immunology; D008545:Melanoma / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; C489519:Mageb3 protein, mouse; D009363:Neoplasm Proteins