| 1. |
- Aquila, Andrew, et al.
(author)
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Time-resolved protein nanocrystallography using an X-ray free-electron laser
- 2012
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In: Optics Express. - 1094-4087. ; 20:3, s. 2706-2716
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Journal article (peer-reviewed)abstract
- We demonstrate the use of an X-ray free electron laser synchronized with an optical pump laser to obtain X-ray diffraction snapshots from the photoactivated states of large membrane protein complexes in the form of nanocrystals flowing in a liquid jet. Light-induced changes of Photosystem I-Ferredoxin co-crystals were observed at time delays of 5 to 10 µs after excitation. The result correlates with the microsecond kinetics of electron transfer from Photosystem I to ferredoxin. The undocking process that follows the electron transfer leads to large rearrangements in the crystals that will terminally lead to the disintegration of the crystals. We describe the experimental setup and obtain the first time-resolved femtosecond serial X-ray crystallography results from an irreversible photo-chemical reaction at the Linac Coherent Light Source. This technique opens the door to time-resolved structural studies of reaction dynamics in biological systems.
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| 2. |
- Johansson, Linda C, 1983, et al.
(author)
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Lipidic phase membrane protein serial femtosecond crystallography.
- 2012
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In: Nature methods. - : Springer Science and Business Media LLC. - 1548-7105 .- 1548-7091. ; 9:3, s. 263-265
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Journal article (peer-reviewed)abstract
- X-ray free electron laser (X-FEL)-based serial femtosecond crystallography is an emerging method with potential to rapidly advance the challenging field of membrane protein structural biology. Here we recorded interpretable diffraction data from micrometer-sized lipidic sponge phase crystals of the Blastochloris viridis photosynthetic reaction center delivered into an X-FEL beam using a sponge phase micro-jet.
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| 3. |
- Johansson, Linda C, 1983, et al.
(author)
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Structure of a photosynthetic reaction centre determined by serial femtosecond crystallography.
- 2013
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In: Nature communications. - : Springer Science and Business Media LLC. - 2041-1723. ; 4
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Journal article (peer-reviewed)abstract
- Serial femtosecond crystallography is an X-ray free-electron-laser-based method with considerable potential to have an impact on challenging problems in structural biology. Here we present X-ray diffraction data recorded from microcrystals of the Blastochloris viridis photosynthetic reaction centre to 2.8Å resolution and determine its serial femtosecond crystallography structure to 3.5Å resolution. Although every microcrystal is exposed to a dose of 33MGy, no signs of X-ray-induced radiation damage are visible in this integral membrane protein structure.
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| 4. |
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| 5. |
- Andersson, Magnus, et al.
(author)
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A proposed time-resolved X-ray scattering approach to track local and global conformational changes in membrane transport proteins
- 2008
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In: Structure. - : Elsevier BV. - 0969-2126 .- 1878-4186. ; 16:1, s. 21-28
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Journal article (peer-reviewed)abstract
- Time-resolved X-ray scattering has emerged as a powerful technique for studying the rapid structural dynamics of small molecules in solution. Membrane-protein-catalyzed transport processes frequently couple large-scale conformational changes of the transporter with local structural changes perturbing the uptake and release of the transported substrate. Using light-driven halide ion transport catalyzed by halorhodopsin as a model system, we combine molecular dynamics simulations with X-ray scattering calculations to demonstrate how small-molecule time-resolved X-ray scattering can be extended to the study of membrane transport processes. In particular, by introducing strongly scattering atoms to label specific positions within the protein and substrate, the technique of time-resolved wide-angle X-ray scattering can reveal both local and global conformational changes. This approach simultaneously enables the direct visualization of global rearrangements and substrate movement, crucial concepts that underpin the alternating access paradigm for membrane transport proteins.
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| 6. |
- Andersson, Magnus, et al.
(author)
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Structural Dynamics of Light-Driven Proton Pumps
- 2009
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In: Structure. - : Elsevier BV. - 0969-2126 .- 1878-4186. ; 17:9, s. 1265-1275
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Journal article (peer-reviewed)abstract
- Bacteriorhodopsin and proteorhodopsin are simple heptahelical proton pumps containing a retinal chromophore covalently bound to helix G via a protonated Schiff base. Following the absorption of a photon, all-trans retinal is isomerized to a 13-cis conformation, initiating a sequence of conformational changes driving vectorial proton transport. In this study we apply time-resolved wide-angle X-ray scattering to visualize in real time the helical motions associated with proton pumping by bacteriorhodopsin and proteorhodopsin. Our results establish that three conformational states are required to describe their photocycles. Significant motions of the cytoplasmic half of helix F and the extracellular half of helix C are observed prior to the primary proton transfer event, which increase in amplitude following proton transfer. These results both simplify the structural description to emerge from intermediate trapping studies of bacteriorhodopsin and reveal shared dynamical principles for proton pumping.
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| 7. |
- Arnlund, David, et al.
(author)
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Visualizing a protein quake with time-resolved X-ray scattering at a free-electron laser
- 2014
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In: Nature Methods. - : Springer Science and Business Media LLC. - 1548-7091 .- 1548-7105. ; 11:9, s. 923-926
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Journal article (peer-reviewed)abstract
- We describe a method to measure ultrafast protein structural changes using time-resolved wide-angle X-ray scattering at an X-ray free-electron laser. We demonstrated this approach using multiphoton excitation of the Blastochloris viridis photosynthetic reaction center, observing an ultrafast global conformational change that arises within picoseconds and precedes the propagation of heat through the protein. This provides direct structural evidence for a 'protein quake': the hypothesis that proteins rapidly dissipate energy through quake-like structural motions.
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| 8. |
- Barty, A., et al.
(author)
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Self-terminating diffraction gates femtosecond X-ray nanocrystallography measurements
- 2012
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In: Nature Photonics. - 1749-4885 .- 1749-4893. ; 6:1, s. 35-40
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Journal article (peer-reviewed)abstract
- X-ray free-electron lasers have enabled new approaches to the structural determination of protein crystals that are too small or radiation-sensitive for conventional analysis1. For sufficiently short pulses, diffraction is collected before significant changes occur to the sample, and it has been predicted that pulses as short as 10 fs may be required to acquire atomic-resolution structural information1, 2, 3, 4. Here, we describe a mechanism unique to ultrafast, ultra-intense X-ray experiments that allows structural information to be collected from crystalline samples using high radiation doses without the requirement for the pulse to terminate before the onset of sample damage. Instead, the diffracted X-rays are gated by a rapid loss of crystalline periodicity, producing apparent pulse lengths significantly shorter than the duration of the incident pulse. The shortest apparent pulse lengths occur at the highest resolution, and our measurements indicate that current X-ray free-electron laser technology5 should enable structural determination from submicrometre protein crystals with atomic resolution.
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| 9. |
- Bertini, Ivano, et al.
(author)
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Search for satellites near (21) Lutetia using OSIRIS/Rosetta images
- 2012
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In: Planetary and Space Science. - : Elsevier BV. - 0032-0633 .- 1873-5088. ; 66:1, s. 64-70
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Journal article (peer-reviewed)abstract
- On 2010 July 10 the ESA Rosetta mission flew by the large asteroid (21) Lutetia. One of the scientific goals of the onboard OSIRIS instrument was the search for satellites of the asteroid, with more than 20 images specifically dedicated to this topic. An observational campaign was devised with a selection of filters and exposure times tailored to maximize the possibility of detecting small companions and determining their bound orbits. Data were analyzed with suitable methods to remove cosmic ray hits and known background objects, in order to search for persistent detections of potential interesting flux sources. We found no unambiguous detections of a satellite larger than similar to 160 m inside the entire sphere of gravitational influence. Our search confirmed the absence of bound companions larger than similar to 30 m inside 20 primary radii. These limits are a factor of similar to 30 smaller than the values reported so far from large ground-based telescopes using adaptive optics and from the Hubble Space Telescope.
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| 10. |
- Björgvinsson, Erling, et al.
(author)
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Prototyping Futures
- 2012
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Reports (other academic/artistic)abstract
- Prototyping Futures gives you a glimpse of what collaborating with academia might look like. Medea and its co-partners share their stories about activities happening at the research centre – projects, methods, tools, and approaches – what challenges lie ahead, and how these can be tackled. Examples of highlighted topics include: What is a living lab and how does it work? What are the visions behind the Connectivity Lab at Medea? And, how can prototyping-methods be used when sketching scenarios for sustainable futures? Other topics are: What is the role of the body when designing technology? What is collaborative media and how can this concept help us understand contemporary media practices? Prototyping Futures also discusses the open-hardware platform Arduino, and the concepts of open data and the Internet of Things, raising questions on how digital media and connected devices can contribute to more sustainable lifestyles, and a better world.
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