Structural Aspects of the Association of FcεRI with Detergent-resistant Membranes*
Abstract
We recently showed that aggregation of the high affinity IgE receptor on mast cells, FcεRI, 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 FcεRI and other receptors to undergo this association. We show that this interaction is not mediated by the β subunit of the receptor or the cytoplasmic tail of the γ subunit, both of which are involved in signaling. Using chimeric receptor constructs, we found that the extracellular segment of the FcεRI α subunit was not sufficient to mediate this association, implicating FcεRI α and/or γ 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 α4 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 α (Tac) on Chinese hamster ovary cells exhibited aggregation-dependent membrane domain association similar to FcεRI. These results provide insights into the structural basis and selectivity of lipid-mediated interactions between certain transmembrane receptors and detergent-resistant membranes.
Footnotes
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↵* This work was supported by Grant AI22449 from the National Institutes of Health.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵‡ Present address: G. W. Hooper Foundation, Box 0552, University of California, San Francisco, CA 94143.
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↵§ To whom correspondence should be addressed. Tel.: 607-255-4095; Fax: 607-255-4137; E-mail: dah24{at}cornell.edu.
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↵2 D. Holowka, unpublished observations.
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↵3 K. A. Field, D. Holowka, and B. Baird, unpublished observations.
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↵4 D. Reczeck and K. Field, unpublished observations.
- Abbreviations:
- PTK
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protein tyrosine kinase
- ITAM
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immunoreceptor tyrosine-based activation motif
- RBL
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rat basophilic leukemia
- DRM
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detergent-resistant membrane
- GPI
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glycosylphosphatidylinositol
- CHO
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Chinese hamster ovary
- IL
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interleukin
- TX-100
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Triton X-100.
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- Received August 18, 1998.
- Revision received October 1, 1998.
- The American Society for Biochemistry and Molecular Biology, Inc.
