100 research outputs found
Three-dimensional structure determination from a single view
The ability to determine the structure of matter in three dimensions has
profoundly advanced our understanding of nature. Traditionally, the most widely
used schemes for 3D structure determination of an object are implemented by
acquiring multiple measurements over various sample orientations, as in the
case of crystallography and tomography (1,2), or by scanning a series of thin
sections through the sample, as in confocal microscopy (3). Here we present a
3D imaging modality, termed ankylography (derived from the Greek words ankylos
meaning 'curved' and graphein meaning 'writing'), which enables complete 3D
structure determination from a single exposure using a monochromatic incident
beam. We demonstrate that when the diffraction pattern of a finite object is
sampled at a sufficiently fine scale on the Ewald sphere, the 3D structure of
the object is determined by the 2D spherical pattern. We confirm the
theoretical analysis by performing 3D numerical reconstructions of a sodium
silicate glass structure at 2 Angstrom resolution and a single poliovirus at 2
- 3 nm resolution from 2D spherical diffraction patterns alone. Using
diffraction data from a soft X-ray laser, we demonstrate that ankylography is
experimentally feasible by obtaining a 3D image of a test object from a single
2D diffraction pattern. This approach of obtaining complete 3D structure
information from a single view is anticipated to find broad applications in the
physical and life sciences. As X-ray free electron lasers (X-FEL) and other
coherent X-ray sources are under rapid development worldwide, ankylography
potentially opens a door to determining the 3D structure of a biological
specimen in a single pulse and allowing for time-resolved 3D structure
determination of disordered materials.Comment: 30 page
Ultrafast structural changes direct the first molecular events of vision
視覚に関わるタンパク質の超高速分子動画 --薄暗いところで光を感じる仕組み--. 京都大学プレスリリース. 2023-03-23.Vision is initiated by the rhodopsin family of light-sensitive G protein-coupled receptors (GPCRs). A photon is absorbed by the 11-cis retinal chromophore of rhodopsin, which isomerizes within 200 femtoseconds to the all-trans conformation, 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 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
Proteorhodopsin Phototrophy Promotes Survival of Marine Bacteria during Starvation
Mutational analysis provides direct evidence for the link between proteorhodopsin light-harvesting and enhanced survival of marine bacteria
Crystal Structure of a Yeast Aquaporin at 1.15 Å Reveals a Novel Gating Mechanism
Atomic-resolution X-ray crystallography, functional analyses, and molecular dynamics simulations suggest a novel mechanism for the regulation of water flux through the yeast Aqy1 water channel
Nanosecond pump–probe device for time-resolved serial femtosecond crystallography developed at SACLA
Membrane protein structural biology using X-ray free electron lasers
Membrane protein structural biology has benefitted tremendously from access to micro-focus crystallography at synchrotron radiation sources. X-ray free electron lasers (XFELs) are linear accelerator driven X-ray sources that deliver a jump in peak X-ray brilliance of nine orders of magnitude and represent a disruptive technology with potential to dramatically change the field. Membrane proteins were amongst the first macromolecules to be studied with XFEL radiation and include proof-of-principle demonstrations of serial femtosecond crystallography (SFX), the observation that XFEL data can deliver damage free crystallographic structures, initial experiments towards recording structural information from 2D arrays of membrane proteins, and time-resolved SFX, time- resolved wide angle X-ray scattering and time-resolved X-ray emission spectroscopy studies. Conversely, serial crystallography methods are now being applied using synchrotron radiation. We believe that a context dependent choice of synchrotron or XFEL radiation will accelerate progress towards novel insights in understanding membrane protein structure and dynamics.ISSN:0959-440XISSN:1879-033
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