Structural and partial amino acid sequence analysis of the human hemopoietic progenitor cell antigen CD34.
Monoclonal antibodies of the CD34 class all recognize a monomeric cell surface antigen of approximately Mr 110,000 which is selectively expressed on human hemopoietic progenitor cells. This structure can be readily surface-labeled with [125I]actoperoxidase and by periodate-[3H]borohydride, but it labels only weakly with [35S]methionine, [35Sl]cysteine, 3H-amino acids, or 3H-mannose, even after prolonged labeling periods. However, the antigen is more efficiently labeled by [3H]glucosamine. Lectin binding studies, sensitivity to certain glycosidases, and gel filtration analysis of glycans released by alkaline hydrolysis indicate that this glycoprotein contains several complex-type N-linked glycans as well as several highly sialylated O-linked glycans. Western blotting experiments show that various CD34 antibodies fail to efficiently detect desialylated and/or de-N-glycosylated forms of the antigen. Experiments involving the use of tunicamycin, together with metabolic labeling studies, strongly suggest that this structure "turns over" very slowly in vivo. The CD34 antigen is not detectably labeled by 32P-phosphate in vivo, nor are immune complexes containing it associated with phosphokinase activity in vitro. Sequential immunoprecipitation and Western blotting studies indicate that this antigen is not a member of the leukosialin/sialophorin family despite the fact that these molecules share several structural similarities. Partial amino acid analysis of highly purified CD34 antigen revealed no significant sequence similarity with any previously described structures.
Sutherland
D R
DR
Oncology Clinic, Toronto General Hospital, Ontario, Canada.
Watt
S M
SM
Dowden
G
G
Karhi
K
K
Baker
M A
MA
Greaves
M F
MF
Smart
J E
JE
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D018952:Antigens, CD34; D000943:Antigens, Differentiation / Q000032:analysis* / Q000096:biosynthesis; D000951:Antigens, Neoplasm / Q000032:analysis; D002846:Chromatography, Affinity; D006031:Glycosylation; D006412:Hematopoietic Stem Cells / Q000276:immunology*; D006801:Humans; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009363:Neoplasm Proteins / Q000032:analysis; D014407:Tumor Cells, Cultured / Q000032:analysis
D000911:Antibodies, Monoclonal; D018952:Antigens, CD34; D000943:Antigens, Differentiation; D000951:Antigens, Neoplasm; D009363:Neoplasm Proteins