Structural aspects of the association of FcepsilonRI with detergent-resistant membranes.
We recently showed that aggregation of the high affinity IgE receptor on mast cells, FcepsilonRI, causes this immunoreceptor to associate rapidly with specialized regions of the plasma membrane, where it is phosphorylated by the tyrosine kinase Lyn. In this study, we further characterize the detergent sensitivity of this association on rat basophilic leukemia-2H3 mast cells, and we compare the capacity of structural variants of FcepsilonRI and other receptors to undergo this association. We show that this interaction is not mediated by the beta subunit of the receptor or the cytoplasmic tail of the gamma subunit, both of which are involved in signaling. Using chimeric receptor constructs, we found that the extracellular segment of the FcepsilonRI alpha subunit was not sufficient to mediate this association, implicating FcepsilonRI alpha and/or gamma transmembrane segments. To determine the specificity of this interaction, we compared the association of several other receptors. Interleukin-1 type I receptors on Chinese hamster ovary cells and alpha4 integrins on rat basophilic leukemia cells showed little or no association with isolated membrane domains, both before and after aggregation on the cells. In contrast, interleukin-2 receptor alpha (Tac) on Chinese hamster ovary cells exhibited aggregation-dependent membrane domain association similar to FcepsilonRI. These results provide insights into the structural basis and selectivity of lipid-mediated interactions between certain transmembrane receptors and detergent-resistant membranes.
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Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York 14853, USA.
Holowka
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Baird
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D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D016466:CHO Cells; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D002499:Centrifugation, Density Gradient; D006224:Cricetinae; D003902:Detergents / Q000494:pharmacology; D039441:Integrin alpha4; D008407:Mast Cells / Q000276:immunology; D051379:Mice; D008854:Microscopy, Electron; D017830:Octoxynol / Q000494:pharmacology; D011487:Protein Conformation; D051381:Rats; D017455:Receptors, IgE / Q000737:chemistry / Q000378:metabolism*; D017472:Receptors, Interleukin-1 / Q000378:metabolism; D015375:Receptors, Interleukin-2 / Q000378:metabolism; D014407:Tumor Cells, Cultured; D019061:src-Family Kinases / Q000378:metabolism
D015703:Antigens, CD; D003902:Detergents; D017455:Receptors, IgE; D017472:Receptors, Interleukin-1; D015375:Receptors, Interleukin-2; D039441:Integrin alpha4; D017830:Octoxynol; C060902:lyn protein-tyrosine kinase; D019061:src-Family Kinases