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Vision is initiated by the rhodopsin family of light-sensitive G protein-coupled receptors (GPCRs) 1 . A photon is absorbed by the 11-cis retinal chromophore of rhodopsin, which isomerizes within 200 femtoseconds to the all-trans conformation 2 , thereby initiating the cellular signal transduction processes that ultimately lead to vision. However, the intramolecular mechanism by which the photoactivated retinal induces the activation events inside rhodopsin remains experimentally unclear. Here we use ultrafast time-resolved crystallography at room temperature 3 to determine how an isomerized twisted all-trans retinal stores the photon energy that is required to initiate the protein conformational changes associated with the formation of the G protein-binding signalling state. The distorted retinal at a 1-ps time delay after photoactivation has pulled away from half of its numerous interactions with its binding pocket, and the excess of the photon energy is released through an anisotropic protein breathing motion in the direction of the extracellular space. Notably, the very early structural motions in the protein side chains of rhodopsin appear in regions that are involved in later stages of the conserved class A GPCR activation mechanism. Our study sheds light on the earliest stages of vision in vertebrates and points to fundamental aspects of the molecular mechanisms of agonist-mediated GPCR activation., (© 2023. The Author(s).)

Mitochondrial energy conversion requires an intricate architecture of the inner mitochondrial membrane 1 . Here we show that a supercomplex containing all four respiratory chain components contributes to membrane curvature induction in ciliates. We report cryo-electron microscopy and cryo-tomography structures of the supercomplex that comprises 150 different proteins and 311 bound lipids, forming a stable 5.8-MDa assembly. Owing to subunit acquisition and extension, complex I associates with a complex IV dimer, generating a wedge-shaped gap that serves as a binding site for complex II. Together with a tilted complex III dimer association, it results in a curved membrane region. Using molecular dynamics simulations, we demonstrate that the divergent supercomplex actively contributes to the membrane curvature induction and tubulation of cristae. Our findings highlight how the evolution of protein subunits of respiratory complexes has led to the I-II-III 2 -IV 2 supercomplex that contributes to the shaping of the bioenergetic membrane, thereby enabling its functional specialization., (© 2023. The Author(s).)

Light-driven sodium pumps actively transport small cations across cellular membranes 1 . These pumps are used by microorganisms to convert light into membrane potential and have become useful optogenetic tools with applications in neuroscience. Although the resting state structures of the prototypical sodium pump Krokinobacter eikastus rhodopsin 2 (KR2) have been solved 2,3 , it is unclear how structural alterations over time allow sodium to be translocated against a concentration gradient. Here, using the Swiss X-ray Free Electron Laser 4 , we have collected serial crystallographic data at ten pump-probe delays from femtoseconds to milliseconds. High-resolution structural snapshots throughout the KR2 photocycle show how retinal isomerization is completed on the femtosecond timescale and changes the local structure of the binding pocket in the early nanoseconds. Subsequent rearrangements and deprotonation of the retinal Schiff base open an electrostatic gate in microseconds. Structural and spectroscopic data, in combination with quantum chemical calculations, indicate that a sodium ion binds transiently close to the retinal within one millisecond. In the last structural intermediate, at 20 milliseconds after activation, we identified a potential second sodium-binding site close to the extracellular exit. These results provide direct molecular insight into the dynamics of active cation transport across biological membranes.

Selective coupling of G protein (heterotrimeric guanine nucleotide-binding protein)-coupled receptors (GPCRs) to specific Gα-protein subtypes is critical to transform extracellular signals, carried by natural ligands and clinical drugs, into cellular responses. At the center of this transduction event lies the formation of a signaling complex between the receptor and G protein. We report the crystal structure of light-sensitive GPCR rhodopsin bound to an engineered mini-G o protein. The conformation of the receptor is identical to all previous structures of active rhodopsin, including the complex with arrestin. Thus, rhodopsin seems to adopt predominantly one thermodynamically stable active conformation, effectively acting like a "structural switch," allowing for maximum efficiency in the visual system. Furthermore, our analysis of the well-defined GPCR-G protein interface suggests that the precise position of the carboxyl-terminal "hook-like" element of the G protein (its four last residues) relative to the TM7/helix 8 (H8) joint of the receptor is a significant determinant in selective G protein activation.

Purpose: A mammographic system has been developed to provide direct radiographic magnification; investigation of image quality and radiation exposure for surveys and aimed exposures were carried out on phantoms and the results compared with two modern screen methods., Material and Method: Image quality was defined by contrast transfer function, resolution of low contrast, contrast of large areas and recognition of low contrast detail, using suitable phantoms. Measurements of entrance surface dose were made with a 1 cm3 flat chamber., Results: For survey examinations, the new mammographic system provided image quality equal to a screen system but using 50% radiation dose. For localised views, the new system used similar radiation dose but provided significantly better demonstration of diagnostic detail., Conclusion: Using a focal spot of 40-120 microns, the new mammographic system provides markedly improved direct radiographic magnification. With reduced radiation exposure at times, there is a significant gain in diagnostic information compared with current standard systems.

Purpose: Since the routine use of storage phosphor systems for mammography has been limited by its inadequate spatial resolution of 5 linepairs/mm, a combination of a magnification mammography technique with storage phosphor plates was investigated to detect microcalcifications., Material and Methods: A new mammography system with a microfocus tube using an anode of 0.05-0.12 mm allowed to obtain survey views of the breast with 1.7x magnification (m), and spot views with 4x magnification. The digital image receptor comprised a high resolution storage phosphor plate. To determine spatial resolution, contrast transfer curves were obtained, and the detection of microcalcifications was investigated by ROC (receiver operating characteristic) analysis., Results: Spatial resolution for digital survey views (m = 1.7) was 8 linepairs/mm and for spot views (m = 4) was 18 linepairs/mm. ROC analysis demonstrated a significantly higher performance of the digital magnification technique compared to the conventional screen-film mammography technique., Conclusions: The limitations of digital mammography with respect to spatial resolution can be overcome by using a high magnification technique.