Expression of the leukocyte-associated sialoglycoprotein CD43 by a colon carcinoma cell line.
The colon adenocarcinoma cell line COLO 205 secretes L-CanAg, a mucin-like glycoprotein carrying the carcinoma-associated sialyl-Lewis a carbohydrate epitope. In an attempt to identify its apoprotein, an NH2-terminal peptide sequence was obtained from purified L-CanAg. In all interpretable positions, this sequence showed 100% identity to the NH2-terminal of human CD43 (leukosialin, sialophorin), a plasma membrane-bound sialoglycoprotein hitherto only identified in leukocytes and other hematopoietic cells. An antiserum against deglycosylated L-CanAg and an anti-CD43 antiserum both immunoprecipitated a 61-kDa band, interpreted as the CD43 precursor, from COLO 205 cells as well as from the known CD43-expressing cell line HL-60. Results from immunoprecipitations following pulse-chase experiments and tunicamycin treatments were in agreement with earlier studies on the CD43 precursor. RNA blot analysis confirmed the expression of CD43 by the COLO 205 cell line, whereas three other colon carcinoma cell lines were negative. The glycosylation-dependent monoclonal antibody Leu-22, which recognizes leukocyte CD43, failed to bind L-CanAg, probably due to its much more extensive glycosylation. We conclude that L-CanAg is the secreted extracellular domain of a novel glycoform of CD43 and that CD43, if expressed in other carcinoma cells, may have escaped notice in studies relying on glycosylation-dependent monoclonal antibodies against leukocyte CD43.
Baeckström
D
D
Department of Medical Biochemistry, University of Göteborg, Sweden.
Zhang
K
K
Asker
N
N
Rüetschi
U
U
Ek
M
M
Hansson
G C
GC
D000230:Adenocarcinoma / Q000737:chemistry*; D000595:Amino Acid Sequence; D000818:Animals; D015703:Antigens, CD; D003110:Colonic Neoplasms / Q000737:chemistry*; D006801:Humans; D051917:Leukosialin; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D011498:Protein Precursors / Q000276:immunology; D012333:RNA, Messenger / Q000032:analysis; D011817:Rabbits; D012795:Sialoglycoproteins / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D014407:Tumor Cells, Cultured
D015703:Antigens, CD; D051917:Leukosialin; D011498:Protein Precursors; D012333:RNA, Messenger; C465014:SPN protein, human; D012795:Sialoglycoproteins