The role of the Fc epsilon receptor in calcium channel opening in rat basophilic leukemia cells.
The role of the Fc epsilon, receptor (Fc epsilon R), isolated from rat basophilic leukemia cells (line RBL-2H3) in antigen induced Ca++ channel opening has been studied by following ion conductance in reconstituted model membranes. Planar bilayers were constructed from lipid vesicles containing the purified Fc epsilon R alone, or together with the cromolyn binding protein (CBP). Changes in conductivity of these bilayers were measured as a monitor for channel activity, following specific aggregation of Fc epsilon R. Antigen-induced, Fc epsilon R mediated channel activity could only be elicited in membranes containing both proteins. This conductance was abrogated upon disaggregating the complexes with a monovalent hapten (epsilon-N-DNP-L-lysine). No channel activity was observed following antigen-induced aggregation of Fc epsilon R if CBP was not present in the bilayer. The single channels recorded were of approximately equal to 2 pS conductance. The open-time values varied significantly with individual experiments and depended on the protein composition of the membrane and the nature of the aggregating agent. These observations strongly indicate that the Fc epsilon R isolated from RBL cells does not form cation (Ca++) channels by itself. Furthermore, in line with earlier reports, the present data suggest that the CBP is responsible for this activity, and that it interacts directly with Fc epsilon R to open channels upon aggregation.
N
N
Mazurek
V
V
Dulic
I
I
Pecht
H G
HG
Schindler
B
B
Rivnay
D000818:Animals; D001491:Basophils / Q000648:ultrastructure; D002118:Calcium / Q000378:metabolism*; D002352:Carrier Proteins / Q000276:immunology; D002460:Cell Line; D002478:Cells, Cultured; D004553:Electric Conductivity; D000939:Epitopes; D007073:Immunoglobulin E / Q000276:immunology; D007473:Ion Channels / Q000276:immunology / Q000378:metabolism*; D007938:Leukemia / Q000378:metabolism*; D008560:Membrane Fluidity; D008565:Membrane Proteins; D051381:Rats; D011961:Receptors, Fc / Q000276:immunology* / Q000302:isolation & purification; D017455:Receptors, IgE
D002352:Carrier Proteins; D000939:Epitopes; D007473:Ion Channels; D008565:Membrane Proteins; D011961:Receptors, Fc; D017455:Receptors, IgE; C039229:cromolyn binding protein, rat; D007073:Immunoglobulin E; D002118:Calcium