Molecular Interactions Leading to the Coadsorption of Surfactant Dodecyltrimethylammonium Bromide and Poly(styrenesulfonate) at the Oil/Water Interface.

The strong synergistic adsorption of mixed polymer/surfactant (P/S) systems at the oil/water interface shows promise for applications such as oil remediation and emulsion stabilization, especially with respect to the formation of tunable mesoscopic multilayers. There is some evidence that a combination of dodecyltrimethylammonium bromide (DTAB) and sodium poly(styrenesulfonate) (PSS) exhibits the adsorption of a secondary P/S layer, though the structure of this layer has long eluded researchers. The focus of this study is to determine whether the DTAB-assisted adsorption of PSS at the oil/water interface occurs as a single layer or with subsequent multilayers. The study presented uses vibrational sum-frequency spectroscopy and interfacial tensiometry to determine the degree of PSS adsorption and orientation of its charged groups relative to the interface at three representative concentrations of DTAB. At low and intermediate DTAB concentrations, a single mixed DTAB/PSS monolayer adsorbs at the oil/water interface. No PSS adsorbs above the system critical micelle concentration. The interfacial charge is found to be similar to that of P/S complexes solvated in the aqueous solution. The surface adsorbate and P/S complexes in the bulk both exhibit a charge inversion at around the same DTAB concentration. This study demonstrates the importance of techniques which can differentiate between coadsorbing species and calls into question current models of P/S adsorption at an oil/water interface.

The heterodimeric assembly of the CD94-NKG2 receptor family and implications for human leukocyte antigen-E recognition.

The CD94-NKG2 receptor family that regulates NK and T cells is unique among the lectin-like receptors encoded within the natural killer cell complex. The function of the CD94-NKG2 receptors is dictated by the pairing of the invariant CD94 polypeptide with specific NKG2 isoforms to form a family of functionally distinct heterodimeric receptors. However, the structural basis for this selective pairing and how they interact with their ligand, HLA-E, is unknown. We describe the 2.5 A resolution crystal structure of CD94-NKG2A in which the mode of dimerization contrasts with that of other homodimeric NK receptors. Despite structural homology between the CD94 and NKG2A subunits, the dimer interface is asymmetric, thereby providing a structural basis for the preferred heterodimeric assembly. Structure-based sequence comparisons of other CD94-NKG2 family members, combined with extensive mutagenesis studies on HLA-E and CD94-NKG2A, allows a model of the interaction between CD94-NKG2A and HLA-E to be established, in which the invariant CD94 chain plays a more dominant role in interacting with HLA-E in comparison to the variable NKG2 chain.

The NMR solution structure of the relaxin (RXFP1) receptor lipoprotein receptor class A module and identification of key residues in the N-terminal region of the module that mediate receptor activation.

The receptors for the peptide hormones relaxin and insulin-like peptide 3 (INSL3) are the leucine-rich repeat-containing G-protein-coupled receptors LGR7 and LGR8 recently renamed as the relaxin family peptide (RXFP) receptors, RXFP1 and RXFP2, respectively. These receptors differ from other LGRs by the addition of an N-terminal low density lipoprotein receptor class A (LDLa) module and are the only human G-protein-coupled receptors to contain such a domain. Recently it was shown that the LDLa module of the RXFP1 and RXFP2 receptors is essential for ligand-stimulated cAMP signaling. The mechanism by which the LDLa module modulates receptor signaling is unknown; however, it represents a unique paradigm in understanding G-protein-coupled receptor signaling. Here we present the structure of the RXFP1 receptor LDLa module determined by solution NMR spectroscopy. The structure is similar to other LDLa modules but shows small differences in side chain orientations and inter-residue packing. Interchange of the module with the second ligand binding domain of the LDL receptor, LB2, results in a receptor that binds relaxin with full affinity but is unable to signal. Furthermore, we demonstrate via structural studies on mutated LDLa modules and functional studies on mutated full-length receptors that a hydrophobic surface within the N-terminal region of the module is essential for activation of RXFP1 receptor signal in response to relaxin stimulation. This study has highlighted the necessity to understand the structural effects of single amino acid mutations on the LDLa module to fully interpret the effects of these mutations on receptor activity.

The human LGR7 low-density lipoprotein class A module requires calcium for structure.

The relaxin and INSL3 receptors, LGR7 and LGR8, are the only human G-protein-coupled receptors to contain a low-density lipoprotein class-A (LDL-A) module. LDL-A modules are well characterized in a variety of diverse biological functions that involve ligand binding to elicit a response. Common features of the LDL-A modules characterized to date are the conservation of three disulfide bonds and the structural arrangement around a bound calcium ion. In this study we recombinantly produce the human LGR7 LDL-A module for NMR studies and demonstrate that calicum is required for the module to form a stable and correctly folded structure.