Two-dimensional analysis of metabolically and cell surface radiolabeled proteins of some human lymphoid and myeloid leukemia cell lines. I. 35S-methionine labeled, lactoperoxidase radioiodinated and 3H-reductively methylated proteins.
Cell surface and cytoplasmic polypeptides of some human hemopoietic cell lines were radiolabeled by three different techniques: 35S-methionine metabolic radiolabeling, lactoperoxidase catalyzed radioiodination and reductive methylation with formaldehyde and tritiated sodium borohydride. Two-dimensional patterns of 35S-methionine labeled proteins revealed more than 500 separated protein spots with essentially similar general aspects of all examined cell lines, sharing major polypeptides such as those with electrophoretic mobilities of major cytoskeletal proteins. 125I-lactoperoxidase radio-iodinated cell surface protein patterns contained about 30 separated protein macromolecules with some marked differences between lymphoid versus myeloid lines and also between individual cell lines. Two-dimensional patterns of tritiated membrane polypeptides consisted of approximately 200 separated protein spots, some prominent of them appeared with electrophoretic mobilities of major cytoskeletal proteins being common to both 35S-methionine and reductive methylation patterns. Two-dimensional patterns of 35S-methionine labeled and cell surface radioiodinated proteins immunoprecipitated by monoclonal antibodies Bra30 and HL39, recognizing MHC class II antigens, appeared as bimolecular complex of larger acidic and smaller basic chains structurally corresponding to the expected two-dimensional patterns of MHC class II antigens.
Duraj
J
J
Chorváth
B
B
Sedlák
J
J
Plesková
I
I
D001345:Autoradiography; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007457:Iodine Radioisotopes; D007784:Lactoperoxidase / Q000494:pharmacology; D007945:Leukemia, Lymphoid / Q000378:metabolism*; D007951:Leukemia, Myeloid / Q000378:metabolism*; D008565:Membrane Proteins / Q000032:analysis*; D008715:Methionine / Q000378:metabolism; D008745:Methylation; D009363:Neoplasm Proteins / Q000032:analysis*; D013462:Sulfur Radioisotopes; D014316:Tritium
D007457:Iodine Radioisotopes; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D013462:Sulfur Radioisotopes; D014316:Tritium; D008715:Methionine; D007784:Lactoperoxidase