Three different NCA species, CGM6/CD67, NCA-95, and NCA-90, are comprised in the major 90 to 100-kDa band of granulocyte NCA detectable upon SDS-polyacrylamide gel electrophoresis.
Human granulocytes express several species of nonspecific cross-reacting antigens (NCA), glycoproteins belonging to the carcinoembryonic antigen (CEA) family. Our previous studies have shown that at least two different NCA of 95 and 90 kDa are contained in the major NCA band of 90 to 100 kDa detectable upon gel electrophoresis of immunoprecipitates obtained from the cell surfaces of granulocytes with polyclonal anti-NCA. In the present study, the 90 to 100-kDa NCA band was found to include one more species of 100 kDa. This component was reactive with an anti-CD67 antibody as well as polyclonal anti-NCA and released from the cell surface with phosphatidylinositol-specific phospholipase C, indicating that the 100-kDa NCA species is CD67. Both antibodies revealed high binding activities with a recombinant protein of CGM6, which has been identified in a leukocyte cDNA library as an NCA gene and found to encode a glycosyl-phosphatidylinositol-anchored heterotypic cell adhesion molecule. Furthermore, the apparent molecular mass of the deglycosylated CD67 (38 kDa) corresponded with that of the CGM6 protein. These results suggest that CD67 is equivalent to the NCA species CGM6.
Department of Biochemistry, School of Medicine, Fukuoka University, Japan.
M
M
Kuroki
Y
Y
Matsuo
T
T
Kinugasa
Y
Y
Matsuoka
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology; D000951:Antigens, Neoplasm; D000954:Antigens, Surface / Q000032:analysis*; D016466:CHO Cells; D002272:Carcinoembryonic Antigen; D015815:Cell Adhesion Molecules; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D000939:Epitopes / Q000032:analysis; D006098:Granulocytes / Q000276:immunology*; D006801:Humans; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000302:isolation & purification; D008970:Molecular Weight; D014162:Transfection
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D000954:Antigens, Surface; D002272:Carcinoembryonic Antigen; D015815:Cell Adhesion Molecules; D000939:Epitopes; D008562:Membrane Glycoproteins