The design of macromolecular crystallography diffraction experiments

Evans, Gwyndaf; Axford, Danny; Owen, Robin L.

doi:10.1107/s0907444911007608

Cited by 50 publications

(40 citation statements)

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“…Diffraction data were collected at 100 K on the microfocus beamline I24 [17] of Diamond Light Source in Didcot, England using a Pilatus 6 M detector (Dectris Ltd, Baden, Switzerland). A dataset of 1800 images covering 360°of rotation was collected from a single crystal.…”

Section: Data Collectionmentioning

confidence: 99%

The crystal structure of Z‐(Aib)₁₀‐OH at 0.65 Å resolution: three complete turns of 3₁₀‐helix

Geßmann

Brückner

Petratos

2015

Journal of Peptide Science

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The synthetic peptide Z-(Aib)10-OH was crystallized from hot methanol by slow evaporation. The crystal used for data collection reflected synchrotron radiation to sub-atomic resolution, where the bonding electron density becomes visible between the non-hydrogen atoms. Crystals belong to the centrosymmetric space group P1. Both molecules in the asymmetric unit form regular 310 -helices. All residues in each molecule possess the same handedness, which is in contrast to all other crystal structure determined to date of longer Aib-homopeptides. These other peptides are C-terminal protected by OtBu or OMe. In these cases, because of the missing ability of the C-terminal protection group to form a hydrogen bond to the residue i-3, the sense of the helix is reversed in the last residue. Here, the C-terminal OH-groups form hydrogen bonds to the residues i-3, in part mediated by water molecules. This makes Z-(Aib)10-OH an Aib-homopeptide with three complete 310-helical turns in spite of the shorter length it has compared with Z-(Aib)11-OtBu, the only homopeptide to date with three complete turns.

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Section: Data Collectionmentioning

confidence: 99%

The crystal structure of Z‐(Aib)₁₀‐OH at 0.65 Å resolution: three complete turns of 3₁₀‐helix

Geßmann

Brückner

Petratos

2015

Journal of Peptide Science

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“…With suitable goniometry, it will be possible to move the structures into position in significantly less than 1 s. This will be essential in order to meet the demands for automated high-throughput macromolecular crystallography experiments when the beam size must be altered to match each crystal sample (Evans et al, 2011).…”

Section: Discussionmentioning

confidence: 99%

Using refractive optics to broaden the focus of an X-ray mirror

Laundy

Sawhney

Dhamgaye

2017

J Synchrotron Radiat

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X-ray mirrors are widely used at synchrotron radiation sources for focusing X-rays into focal spots of size less than 1 mm. The ability of the beamline optics to change the size of this spot over a range up to tens of micrometres can be an advantage for many experiments such as X-ray microprobe and X-ray diffraction from micrometre-scale crystals. It is a requirement that the beam size change should be reproducible and it is often essential that the change should be rapid, for example taking less than 1 s, in order to allow high data collection rates at modern X-ray sources. In order to provide a controlled broadening of the focused spot of an X-ray mirror, a series of refractive optical elements have been fabricated and installed immediately before the mirror. By translation, a new refractive element is moved into the X-ray beam allowing a variation in the size of the focal spot in the focusing direction. Measurements using a set of prefabricated refractive structures with a test mirror showed that the focused beam size could be varied from less than 1 mm to over 10 mm for X-rays in the energy range 10-20 keV. As the optics is in-line with the X-ray beam, there is no effect on the centroid position of the focus. Accurate positioning of the refractive optics ensures reproducibility in the focused beam profile and no additional re-alignment of the optics is required.

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“…By adjusting the beam focus to the size of the crystal, the background scattering from the buffer surrounding the crystal can be minimized and signal to noise maximised (Evans et al , 2011). Where the beamline’s native focus is larger than the sample, the beam size can be adjusted at the expense of flux through the use of slits or pinhole apertures (as demonstrated for example at GM/CA, APS (Fischetti et al , 2009) or offered as part of the functionality of the MD2 microdiffractometer).…”

Section: Endstation and Experimentsmentioning

confidence: 99%

Current advances in synchrotron radiation instrumentation for MX experiments

Owen

Juanhuix

Fuchs

2016

Archives of Biochemistry and Biophysics

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Following pioneering work 40 years ago, synchrotron beamlines dedicated to macromolecular crystallography (MX) have improved in almost every aspect as instrumentation has evolved. Beam sizes and crystal dimensions are now on the single micron scale while data can be collected from proteins with molecular weights over 10 MDa and from crystals with unit cell dimensions over 1000 Å. Furthermore it is possible to collect a complete data set in seconds, and obtain the resulting structure in minutes. The impact of MX synchrotron beamlines and their evolution is reflected in their scientific output, and MX is now the method of choice for a variety of aims from ligand binding to structure determination of membrane proteins, viruses and ribosomes, resulting in a much deeper understanding of the machinery of life. A main driving force of beamline evolution have been advances in almost every aspect of the instrumentation comprising a synchrotron beamline. In this review we aim to provide an overview of the current status of instrumentation at modern MX experiments. The most critical optical components are discussed, as are aspects of endstation design, sample delivery, visualization and positioning, the sample environment, beam shaping, detectors and data acquisition and processing.

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The design of macromolecular crystallography diffraction experiments

Cited by 50 publications

References 50 publications

The crystal structure of Z‐(Aib)₁₀‐OH at 0.65 Å resolution: three complete turns of 3₁₀‐helix

The crystal structure of Z‐(Aib)₁₀‐OH at 0.65 Å resolution: three complete turns of 3₁₀‐helix

Using refractive optics to broaden the focus of an X-ray mirror

Current advances in synchrotron radiation instrumentation for MX experiments

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The design of macromolecular crystallography diffraction experiments

Cited by 50 publications

References 50 publications

The crystal structure of Z‐(Aib)10‐OH at 0.65 Å resolution: three complete turns of 310‐helix

The crystal structure of Z‐(Aib)10‐OH at 0.65 Å resolution: three complete turns of 310‐helix

Using refractive optics to broaden the focus of an X-ray mirror

Current advances in synchrotron radiation instrumentation for MX experiments

Contact Info

Product

Resources

About

The crystal structure of Z‐(Aib)₁₀‐OH at 0.65 Å resolution: three complete turns of 3₁₀‐helix

The crystal structure of Z‐(Aib)₁₀‐OH at 0.65 Å resolution: three complete turns of 3₁₀‐helix