GFAP and nuclear lamins share an epitope recognized by monoclonal antibody J1-31.
Monoclonal antibody J1-31 was raised against plaque materials taken from brains of patients who had suffered from multiple sclerosis (MS). Preliminary characterization of the antigen revealed it to be a protein of M(w) 68-70 kDa with both a cytoplasmic and nuclear localization. Here we report the results of isolation and peptide sequencing of the antigen from human brains, and immunocytochemical analysis of the antigen in F98 glioma cells. Purification and peptide sequencing indicate that the antibody recognizes a form of glial fibrillary acidic protein, possibly a phosphorylated variant. However, confocal immunocytochemistry and western analysis of F98 glioma cells raise the possibility that it also recognizes a phosphorylated epitope found in nuclear lamins. Analysis of the expression of the J1-31 epitope in F98 cells with respect to time in culture, cell density, and DNA synthesis showed a developmental relationship: cells that were engaged in rapid growth and DNA synthesis exhibited strong J1-31 staining in nuclei, whereas quiescent cells did not. We conclude that mAB J1-31 remains a useful antibody for studying multiple sclerosis, and is likely to prove useful in studies of the dynamics of nuclear lamins, particularly in models for wound-healing.
García
Dana M
DM
Department of Biology, Southwest Texas State University, San Marcos, TX 78666, USA. dana_garcia@swt.edu
Weigum
Shannon E
SE
Koke
Joseph R
JR
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001921:Brain / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D005904:Glial Fibrillary Acidic Protein / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D034904:Lamin Type A / Q000276:immunology / Q000378:metabolism; D034921:Lamin Type B / Q000276:immunology / Q000378:metabolism; D034882:Lamins / Q000276:immunology / Q000378:metabolism*; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D009103:Multiple Sclerosis / Q000473:pathology; D010446:Peptide Fragments / Q000737:chemistry; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000939:Epitopes; D005904:Glial Fibrillary Acidic Protein; D034904:Lamin Type A; D034921:Lamin Type B; D034882:Lamins; D010446:Peptide Fragments; C440308:lamin C