Ligand-induced association of surface immunoglobulin with the detergent insoluble cytoskeleton may involve alpha-actinin.
B cell surface immunoglobulin (SIg) plays an important role in antigen recognition and cellular activation. Cross-linking of SIg by bivalent antibody converts it into a detergent insoluble state. The resultant SIg may be partially solubilized by incubating the detergent insoluble cytoskeleton in buffers that convert F actin to G actin. Immunoprecipitation of SIg from the detergent soluble fraction of [35S]methionine-labeled B cells results in the co-isolation of 112 kDa, 42 kDa, (actin), and three additional proteins in the 70- to 73-kDa molecular mass range, isoelectric point 4.8 to 5.6. Analysis of anti-Ig immunoprecipitates made after preclearing with anti-alpha-actinin showed complete depletion of the 112-kDa protein, suggesting that the 112-kDa protein is immunologically related to alpha-actinin. These immunoprecipitates also showed partial depletion of 70- to 73-kDa proteins and mu and delta heavy chains. After treatment of a rat B cells with anti-Ig, much of the Ig-associated 112-kDa protein and 70- to 73-kDa proteins became detergent insoluble, concomitant with most of the SIg. The migration of the SIg-associated 112-kDa and 70- to 73-kDa proteins from the detergent soluble fraction to the detergent insoluble fraction after ligand treatment, suggests that these proteins might be involved in linking SIg to the underlying cytoskeleton and could be involved in the transmission of mitogenic signals.
Gupta
S K
SK
Department of Pathology, University of Massachusetts Medical Center, Worcester 01605.
Woda
B A
BA
D000185:Actinin / Q000378:metabolism*; D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism* / Q000648:ultrastructure; D011232:Chemical Precipitation; D003599:Cytoskeleton / Q000378:metabolism*; D007140:Immunoglobulin Fab Fragments / Q000378:metabolism*; D066298:In Vitro Techniques; D007526:Isoelectric Point; D008970:Molecular Weight; D051381:Rats; D011947:Receptors, Antigen, B-Cell / Q000378:metabolism*
D007140:Immunoglobulin Fab Fragments; D011947:Receptors, Antigen, B-Cell; D000185:Actinin