Georg Weidenspointner

Femtosecond x-ray protein nanocrystallography

X-ray crystallography provides the vast majority of macromolecular structures, but the success of the method relies on growing crystals of sufficient size. In conventional measurements, the necessary increase in X-ray dose to record data from crystals that are too small leads to extensive damage before a diffraction signal can be recorded. It is particularly challenging to obtain large, well-diffracting crystals of membrane proteins, for which fewer than 300 unique structures have been determined despite their importance in all living cells. Here we present a method for structure determination where single-crystal X-ray diffraction ‘snapshots’ are collected from a fully hydrated stream of nanocrystals using femtosecond pulses from a hard-X-ray free-electron laser, the Linac Coherent Light Source. We prove this concept with nanocrystals of photosystem I, one of the largest membrane protein complexes. More than 3,000,000 diffraction patterns were collected in this study, and a three-dimensional data set was assembled from individual photosystem I nanocrystals (∼200 nm to 2 μm in size). We mitigate the problem of radiation damage in crystallography by using pulses briefer than the timescale of most damage processes. This offers a new approach to structure determination of macromolecules that do not yield crystals of sufficient size for studies using conventional radiation sources or are particularly sensitive to radiation damage.

Single mimivirus particles intercepted and imaged with an X-ray laser

X-ray lasers offer new capabilities in understanding the structure of biological systems, complex materials and matter under extreme conditions. Very short and extremely bright, coherent X-ray pulses can be used to outrun key damage processes and obtain a single diffraction pattern from a large macromolecule, a virus or a cell before the sample explodes and turns into plasma. The continuous diffraction pattern of non-crystalline objects permits oversampling and direct phase retrieval. Here we show that high-quality diffraction data can be obtained with a single X-ray pulse from a non-crystalline biological sample, a single mimivirus particle, which was injected into the pulsed beam of a hard-X-ray free-electron laser, the Linac Coherent Light Source. Calculations indicate that the energy deposited into the virus by the pulse heated the particle to over 100,000 K after the pulse had left the sample. The reconstructed exit wavefront (image) yielded 32-nm full-period resolution in a single exposure and showed no measurable damage. The reconstruction indicates inhomogeneous arrangement of dense material inside the virion. We expect that significantly higher resolutions will be achieved in such experiments with shorter and brighter photon pulses focused to a smaller area. The resolution in such experiments can be further extended for samples available in multiple identical copies.

SPI: The spectrometer aboard INTEGRAL

SPI is a high spectral resolution gamma-ray telescope on board the ESA mission INTEGRAL (International Gamma Ray Astrophysics Laboratory). It consists of an array of 19 closely packed germanium detectors surrounded by an active anticoincidence shield of BGO. The imaging capabilities of the instrument are obtained with a tungsten coded aperture mask located 1.7 m from the Ge array. The fully coded field-of-view is 16degrees, the partially coded field of view amounts to 31degrees, and the angular resolution is 2.5degrees. The energy range extends from 20 keV to 8 MeV with a typical energy resolution of 2.5 keV at 1.3 MeV. Here we present the general concept of the instrument followed by a brief description of each of the main subsystems. INTEGRAL was successfully launched in October 2002 and SPI is functioning extremely well.

Early SPI/INTEGRAL measurements of 511 keV line emission from the 4th quadrant of the Galaxy

We report the first measurements of the 511 keV line emission from the Galactic Centre (GC) region performed with the spectrometer SPI on the space observatory INTEGRAL (International Gamma-Ray Astrophysics Laboratory). Taking into account the range of spatial distribution models which are consistent with the data, we derive a flux of 9:9 +4:7 2:1 10 4 ph cm 2 s 1 and an intrinsic line width of 2:95 +0:45 0:51 keV (FWHM). The results are consistent with other high-spectroscopy measurements, though the width is found to be at the upper bound of previously reported values.We report the first measurements of the 511 keV line emission from the Galactic Centre (GC) region performed with the spectrometer SPI on the space observatory INTEGRAL (International Gamma-Ray Astrophysics Laboratory). Taking into account the range of spatial distribution models which are consistent with the data, we derive a flux of

Time-resolved protein nanocrystallography using an X-ray free-electron laser

9.9^{+4.7}_{-2.1} \times 10^{-4}

An asymmetric distribution of positrons in the galactic disk revealed by big gamma-rays

ph cm

In vivo protein crystallization opens new routes in structural biology

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Lipidic phase membrane protein serial femtosecond crystallography.

s

Fractal morphology, imaging and mass spectrometry of single aerosol particles in flight

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X-Ray Diffraction from Isolated and Strongly Aligned Gas-Phase Molecules with a Free-Electron Laser

and an intrinsic line width of