Membrane IgM: interactions with the cortical cytoskeleton in the human lymphoblastoid cell line WiL2.
Cell-surface IgM (antigen receptor) sediments with the membrane fraction following osmotic lysis and homogenization of cells of the human lymphoblastoid cell line WiL2. In nonreducing buffers, SDS PAGE analysis of membrane pellets demonstrates that "native" membrane IgM exists as a dimer. In contrast to osmotic lysis, lysis of cells with the nonionic detergent Triton X-100 releases approximately 90% of the membrane-bound IgM into the supernatant; approximately 10% of the IgM pellets with the cytoskeletal fraction on centrifugation. Ligand challenge with either mu-chain-specific antibodies or concanavalin A induces a change in the state of membrane IgM making it refractory to detergent extraction, such that 43% of the IgM pellets during centrifugation. This ligand-induced retention of IgM is significantly diminished by the microfilament-disrupting agent cytochalasin D, whereas pretreatment of cells with sodium azide or colchicine results in no significant change in the percentage of membrane IgM retained by Triton X-100 residues. These results indicate that retention of IgM involves an association with the cortical actin-based cytoskeleton. Investigation of the structural basis for ligand-induced Triton X-100 retention of membrane IgM by using ferritin-conjugated antibodies, myosin subfragment S1, and stereo-imaging electron microscopy has revealed linkages between ligand-receptor (antigen-IgM) complexes and elements of the cortical actin-based cytoskeleton.
Salisbury
J L
JL
Department of Developmental Genetics and Anatomy, School of Medicine, Case Western Reserve University, Cleveland.
Baron
A T
AT
Keller
G A
GA
Skiest
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D
D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology / Q000648:ultrastructure; D003599:Cytoskeleton / Q000276:immunology* / Q000648:ultrastructure; D006801:Humans; D007075:Immunoglobulin M / Q000302:isolation & purification*; D007945:Leukemia, Lymphoid; D008854:Microscopy, Electron; D011947:Receptors, Antigen, B-Cell / Q000302:isolation & purification*
D007075:Immunoglobulin M; D011947:Receptors, Antigen, B-Cell