Phosphorylation of the B1 (CD20) molecule by normal and malignant human B lymphocytes.
The B1 molecule (CD20) is a phosphoprotein found only on B lymphocytes. Multiple isoforms of the B1 molecule are expressed with Mr of 33,000, B1(33) and Mr of 34,500-36,000, B1(35). In this study it was found that nonproliferating B cells did not incorporate 32PO4 into B1 although phosphorylated class I histocompatibility molecules were easily detected. In contrast B1 isolated from proliferating or malignant B cells or B cell lines was heavily phosphorylated. Cross-linking B1 on the cell surface by antibody resulted in enhanced phosphorylation of B1 as did exposure to phorbol esters, and the membrane permeable diacylglycerol analog 1,2,-dioctanoylglyceron. B1(33) and B1(35) produced identical peptide maps following limited proteinase digestion. However, B1(35) contained both phosphoserine and phosphothreonine, while B1(33) only contained phosphoserine. In addition alkaline phosphatase was able to remove the phosphate residue(s) that resulted in generation of the B1(35) form of B1 but was unable to remove the phosphorylation of B1(33). These results suggest that phosphorylation of B1 molecules is associated with proliferation and that the different Mr forms of B1 result from the phosphorylation of B1 at different sites. Also, the finding that antibody binding to B1 generated a transmembrane signal may explain why antibody binding to B1 alters B cell function.
Tedder
T F
TF
Division of Tumor Immunology, Dana-Farber Cancer Institute, Boston, Massachusetts 02115.
Schlossman
S F
SF
D000906:Antibodies / Q000502:physiology; D000941:Antigens / Q000276:immunology; D018951:Antigens, CD20; D000944:Antigens, Differentiation, B-Lymphocyte / Q000276:immunology / Q000378:metabolism*; D001402:B-Lymphocytes / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007163:Immunosorbent Techniques; D007700:Kinetics; D007945:Leukemia, Lymphoid / Q000378:metabolism*; D008970:Molecular Weight; D010446:Peptide Fragments / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D010768:Phosphoserine / Q000378:metabolism; D010769:Phosphothreonine / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D000906:Antibodies; D000941:Antigens; D018951:Antigens, CD20; D000944:Antigens, Differentiation, B-Lymphocyte; D010446:Peptide Fragments; D010710:Phosphates; D010750:Phosphoproteins; D010769:Phosphothreonine; D010768:Phosphoserine; D013755:Tetradecanoylphorbol Acetate