Transmembrane features governing Fc receptor CD16A assembly with CD16A signaling adaptor molecules

Many activating immunoreceptors associate with signaling adaptor molecules like Fc epsilon R1 gamma or CD247. Fc epsilon R1 gamma and CD247 share high sequence homology and form disulphide-linked homodimers that contain a pair of acidic aspartic acid residues in their transmembrane (TM) domains that mediate assembly, via interaction with an arginine residue at a similar register to these aspartic acids, with the activating immunoreceptors. However, this model cannot hold true for receptors like CD16A, whose TM domains do not contain basic residues. We have carried out an extensive site-directed mutagenesis analysis of the CD16A receptor complex and now report that the association of receptor with the signaling adaptor depends on a network of polar and aromatic residues along the length of the TM domain. Molecular modeling indicates that CD16A TM residues F-202, D-205, and T-206 form the core of the membrane-embedded trimeric interface by establishing highly favorable contacts to the signaling modules through rearrangement of a hydrogen bond network previously identified in the CD247 TM dimer solution NMR structure. Strikingly, the amino acid D-205 also regulates the turnover and surface expression of CD16A in the absence of Fc epsilon R1 gamma or CD247. Modeling studies indicate that similar features underlie the association of other activating immune receptors, including CD64 and Fc epsilon R1 alpha, with signaling adaptor molecules, and we confirm experimentally that equivalent F, D, and T residues in the TM domain of FceR1a markedly influence the biology of this receptor and its association with Fc epsilon R1 gamma.