Evolutionary action and structural basis of the allosteric switch controlling β(2)AR functional selectivity

Functional selectivity of G-protein-coupled receptors is believed to originate from ligand-specific conformations that activate only subsets of signaling effectors. In this study, to identify molecular motifs playing important roles in transducing ligand binding into distinct signaling responses, we combined in silico evolutionary lineage analysis and structure-guided site-directed mutagenesis with large-scale functional signaling characterization and non-negative matrix factorization clustering of signaling profiles. Clustering based on the signaling profiles of 28 variants of the β(2)-adrenergic receptor reveals three clearly distinct phenotypical clusters, showing selective impairments of either the Gi or βarrestin/endocytosis pathways with no effect on Gs activation. Robustness of the results is confirmed using simulation-based error propagation. The structural changes resulting from functionally biasing mutations centered around the DRY, NPxxY, and PIF motifs, selectively linking these micro-switches to unique signaling profiles. Our data identify different receptor regions that are important for the stabilization of distinct conformations underlying functional selectivity

Ultrashort, MeV-scale laser-plasma positron source for positron annihilation lifetime spectroscopy

Sub-micron defects represent a well-known fundamental problem in manufacturing since they can significantly affect performance and lifetime of virtually any high-value component. Positron annihilation lifetime spectroscopy is arguably the only established method capable of detecting defects down to the sub-nanometer scale but, to date, it only works for surface studies, and with limited resolution. Here, we experimentally and numerically show that laser-driven systems can overcome these well-known limitations, by generating ultra-short positron beams with a kinetic energy tuneable from 500 keV up to 2 MeV and a number of positrons per shot in a 50 keV energy slice \color{black} of the order of $10^3$. Numerical simulations of the expected performance of a typical mJ-scale kHz laser demonstrate the possibility of generating MeV-scale narrow-band and ultra-short positron beams with a flux exceeding $10^5$ positrons/s, of interest for fast volumetric scanning of materials at high resolution.Comment: Submitted to PRA