Structure-function relationships in the mast cell high affinity receptor for IgE. Role of the cytoplasmic domains and of the beta subunit.
To define functionally critical regions of the high affinity receptor for IgE (Fc epsilon RI), we stably transfected P815 cells with mutated cDNAs coding for subunits with truncated cytoplasmic domains (CD). In addition, to examine further the role of the beta subunit, stable transfectants expressing chimeric Fc epsilon RI without beta subunits were generated. Transfectants were tested for receptor-mediated changes in intracellular Ca2+, for stimulated hydrolysis of phosphoinositides, and for protein tyrosine phosphorylation. In all cases these biochemical signals were affected coordinately, suggesting that they are coupled, possibly in a single pathway. Truncation of the alpha subunit or of the NH2-terminal CD of the beta subunit had no effect, but Fc epsilon RIs with beta subunits missing the COOH-terminal CD were inactive. Interestingly, receptors in cells transfected only with human Fc epsilon RI(alpha) (which utilize the gamma chains endogenously synthesized by the P815 cells but which contain no beta subunits) responded normally. Therefore, the beta subunit influences the functions studied but is not essential. Although structural analysis excluded a straightforward mechanism, truncation of the CD of the gamma chain led to loss of signaling.
Alber
G
G
National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institute of Health, Bethesda, Maryland 20892.
Miller
L
L
Jelsema
C L
CL
Varin-Blank
N
N
Metzger
H
H
D000818:Animals; D000911:Antibodies, Monoclonal; D000944:Antigens, Differentiation, B-Lymphocyte / Q000235:genetics / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D003593:Cytoplasm / Q000276:immunology; D005822:Genetic Vectors; D007073:Immunoglobulin E / Q000378:metabolism*; D007700:Kinetics; D046911:Macromolecular Substances; D008407:Mast Cells / Q000276:immunology*; D051379:Mice; D008970:Molecular Weight; D016297:Mutagenesis, Site-Directed; D010716:Phosphatidylinositols / Q000378:metabolism; D010766:Phosphorylation; D010954:Plasmacytoma; D011961:Receptors, Fc / Q000235:genetics / Q000378:metabolism*; D017455:Receptors, IgE; D014162:Transfection
D000911:Antibodies, Monoclonal; D000944:Antigens, Differentiation, B-Lymphocyte; D046911:Macromolecular Substances; D010716:Phosphatidylinositols; D011961:Receptors, Fc; D017455:Receptors, IgE; D007073:Immunoglobulin E; D002118:Calcium