The Journal of Immunology, Vol 151, Issue 5 2760-2774, Copyright © 1993 by American Association of Immunologists

ARTICLES

Effects of subunit mutation on the localization to coated pits and internalization of cross-linked IgE-receptor complexes

SY Mao, JR Pfeiffer, JM Oliver and H Metzger
Section on Chemical Immunology, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, MD 20892.

IgE receptors of mast cells, Fc epsilon RI, localize to coated pits and internalize after cross-linking. We investigated whether any one of the receptor's four distinctive cytoplasmic domains regulates these phenomena. COS cells, which lack Fc epsilon RI entirely, and P815 mouse mastocytoma cells that lack the alpha and beta subunits of the tetrameric Fc epsilon RI (alpha beta gamma 2), were transfected with wild-type, incomplete, or variant Fc epsilon RI. IgE-receptor complexes were observed by electron microscopy. Before cross-linking with anti- IgE gold particles, receptors were not preferentially localized to coated pits, which occupy approximately 1% of the cell surface. After cross-linking, up to 10 to 20% of the wild-type and most other receptor variants were in coated pits in transfected P815 cells at any one time. beta-less variants localized normally but, surprisingly, receptors containing a variant beta subunit showed reduced localization. "Receptors" consisting simply of the lipid-anchored ectodomains of the human alpha subunit failed to localize to coated pits. In general, cross-linked receptors that localized to coated pits were progressively internalized, whereas receptors that failed to accumulate in coated pits were not. We conclude that no single cytoplasmic domain of the Fc epsilon RI uniquely controls its ligand-induced localization to coated pits and internalization.

This article has been cited by other articles:

				C. Oliver, A. Fujimura, A. M. M. Silveira e Souza, R. Orlandini de Castro, R. P. Siraganian, and M. C. Jamur Mast Cell-specific Gangliosides and Fc{varepsilon}RI Follow the Same Endocytic Pathway From Lipid Rafts in RBL-2H3 Cells J. Histochem. Cytochem., April 1, 2007; 55(4): 315 - 325. [Abstract] [Full Text] [PDF]

				J.-S. Shin, C. P. Shelburne, C. Jin, E. A. LeFurgey, and S. N. Abraham Harboring of Particulate Allergens within Secretory Compartments by Mast Cells following IgE/Fc{epsilon}RI-Lipid Raft-Mediated Phagocytosis J. Immunol., November 1, 2006; 177(9): 5791 - 5800. [Abstract] [Full Text] [PDF]

				V. Lam, J. Kalesnikoff, C. W. K. Lee, V. Hernandez-Hansen, B. S. Wilson, J. M. Oliver, and G. Krystal IgE alone stimulates mast cell adhesion to fibronectin via pathways similar to those used by IgE + antigen but distinct from those used by Steel factor Blood, August 15, 2003; 102(4): 1405 - 1413. [Abstract] [Full Text] [PDF]

				B. S. Wilson, J. R. Pfeiffer, and J. M. Oliver Observing Fc{varepsilon}ri Signaling from the Inside of the Mast Cell Membrane J. Cell Biol., May 29, 2000; 149(5): 1131 - 1142. [Abstract] [Full Text] [PDF]

				F Santini, R. Penn, A. Gagnon, J. Benovic, and J. Keen Selective recruitment of arrestin-3 to clathrin coated pits upon stimulation of G protein-coupled receptors J. Cell Sci., January 7, 2000; 113(13): 2463 - 2470. [Abstract] [PDF]

				S. A. Barker, K. K. Caldwell, J. R. Pfeiffer, and B. S. Wilson Wortmannin-Sensitive Phosphorylation, Translocation, and Activation of PLCgamma 1, but Not PLCgamma 2, in Antigen-stimulated RBL-2H3 Mast Cells Mol. Biol. Cell, February 1, 1998; 9(2): 483 - 496. [Abstract] [Full Text]

				K Xu, R. Williams, D Holowka, and B Baird Stimulated release of fluorescently labeled IgE fragments that efficiently accumulate in secretory granules after endocytosis in RBL-2H3 mast cells J. Cell Sci., January 8, 1998; 111(16): 2385 - 2396. [Abstract] [PDF]