Purification of a tumor-specific PNA-binding glycoprotein, gp200, from a human embryonal carcinoma cell line.
A 200-kDa peanut agglutinin (PNA)-binding glycoprotein, gp200, has been purified and partially characterized from the human embryonal carcinoma cell line, HT-E (833k). Tissue distribution analysis of this molecule by lectin blotting with PNA of detergent-extracted proteins from human cell lines and tissues demonstrated expression limited to nonseminomatous germ cell tumors. The 200-kDa protein was purified with lectin affinity and gel filtration chromatography. Purification to apparent homogeneity was demonstrated by one- and two-dimensional gel electrophoresis. Characterization of gp200 revealed it to be a surface integral membrane glycoprotein; however, gp200 could also be purified from the culture media of EC cells, suggesting gp200 has an extracellular role. The carbohydrate groups of gp200 are N-linked and partially sialylated and contain terminal galactose residues. These initial studies suggest that the PNA-defined glycoprotein, gp200, is a candidate for a nonseminomatous germ cell tumor marker.
Schopperle
W M
WM
Division of Cellular Genetics, Charles A. Dana Research Institute, Beth Israel Hospital, Boston, Massachusetts 02215.
Armant
D R
DR
DeWolf
W C
WC
D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006023:Glycoproteins / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D008297:Male; D008970:Molecular Weight; D009928:Organ Specificity; D011975:Receptors, Mitogen / Q000302:isolation & purification* / Q000378:metabolism; D013724:Teratoma; D014407:Tumor Cells, Cultured
D006023:Glycoproteins; D011975:Receptors, Mitogen; C038223:peanut agglutinin receptor