title: The first transmembrane region of the beta-chain stabilizes the tetrameric Fc epsilon RI complex.
abstract: The family of activating immune receptors stabilizes via the 3-helix assembly principle. A charged basic transmembrane residue interacts with two charged acidic transmembrane residues and forms a 3-helix interface to stabilize receptor complexes in the lipid bilayer. One family member, the high affinity receptor for IgE, Fc epsilon RI, is a key regulator of immediate allergic responses. Tetrameric Fc epsilon RI consists of the IgE-binding alpha-chain, the multimembrane-spanning beta-chain and a dimer of the gamma-subunit (Fc epsilon R gamma). Comparative analysis of these seven transmembrane regions indicates that Fc epsilon RI does not meet the charge requirements for the 3-helix assembly mechanism. We performed alanine mutagenesis to show that the only basic amino acid in the transmembrane regions, beta K97, is not involved in Fc epsilon RI stabilization or surface upregulation, a hallmark function of the beta-chain. Even a beta K97E mutant is functional despite four negatively charged acidic amino acids in the transmembrane regions. Using truncation mutants, we demonstrate that the first uncharged transmembrane domain of the beta-chain contains the interface for receptor stabilization. In vitro translation experiments depict the first transmembrane region as the internal signal peptide of the beta-chain. We also show that this beta-chain domain can function as a cleavable signal peptide when used as a leader peptide for a Type I protein. Our results provide evidence that tetrameric Fc epsilon RI does not assemble according to the 3-helix assembly principle. We conclude that receptors formed with multispanning proteins use different mechanisms of shielding transmembrane charged amino acids.
authors.affiliation: Department of Medicine, Division of Gastroenterology and Nutrition, Children's Hospital Boston, 300 Longwood Avenue, EN720, Boston, MA 02115, USA.
authors.forename: Theresa E
authors.initials: TE
authors.lastname: Singleton
authors.forename: Barbara
authors.initials: B
authors.lastname: Platzer
authors.forename: Eleonora
authors.initials: E
authors.lastname: Dehlink
authors.forename: Edda
authors.initials: E
authors.lastname: Fiebiger
mesh_terms: D000595:Amino Acid Sequence; D002460:Cell Line; D006801:Humans; D008051:Lipid Bilayers / Q000737:chemistry*; D008969:Molecular Sequence Data; D009154:Mutation; D011487:Protein Conformation; D055503:Protein Multimerization; D055550:Protein Stability; D017434:Protein Structure, Tertiary; D017455:Receptors, IgE / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics
chemical_list: D008051:Lipid Bilayers; D017455:Receptors, IgE