Crystal transfer experiments carried out with crystals of tryanosomal triosephosphate isomerase (TIM)

Wierenga, R.K.; Zeelen, J.P.; Noble, M.E.M.

doi:10.1016/0022-0248(92)90250-m

Cited by 11 publications

(6 citation statements)

References 18 publications

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“…However, we were unable to identify alternative crystallization conditions which prompted us to consider crystal transfers to low-salt stabilizing solutions containing either PEG 3350 (polyethylene glycol of average molecular weight 3350) or 2-methyl-2,4-pentanediol (MPD) for soaking experiments. 22 Both polyoles yielded conditions in which the transferred ATAD2 crystals remained stabled. Combining the crystal transfer with Kac soaking yielded the acetyl-lysine complex structure, in which the ligand was well dened by electron density (Fig.…”

Section: Resultsmentioning

confidence: 99%

Structure-based approaches towards identification of fragments for the low-druggability ATAD2 bromodomain

et al. 2014

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Section: Resultsmentioning

confidence: 99%

Structure-based approaches towards identification of fragments for the low-druggability ATAD2 bromodomain

et al. 2014

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“…Limiting this flow can help to preserve crystal order and yield a lower mosaicity. Some strategies to reduce this flow are to equilibrate using serial soaks (Garman, 1999), equilibrate via vapor diffusion (David & Burley, 1991;Wierenga et al, 1992) and to adjust the composition of the cryobuffer so the osmotic pressures are identical (David, 1999). In all cases, an important question to consider is the time necessary for full equilibration with the cryosolution.…”

Section: Cooling: Equilibrating the Crystalmentioning

confidence: 99%

Progress in rational methods of cryoprotection in macromolecular crystallography

Alcorn

Juers

2010

Acta Crystallogr D Biol Crystallogr

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Cryogenic cooling of macromolecular crystals is commonly used for X-ray data collection both to reduce crystal damage from radiation and to gather functional information by cryogenically trapping intermediates. However, the cooling process can damage the crystals. Limiting cooling-induced crystal damage often requires cryoprotection strategies, which can involve substantial screening of solution conditions and cooling protocols. Here, recent developments directed towards rational methods for cryoprotection are described. Crystal damage is described in the context of the temperature response of the crystal as a thermodynamic system. As such, the internal and external parts of the crystal typically have different cryoprotection requirements. A key physical parameter, the thermal contraction, of 26 different cryoprotective solutions was measured between 294 and 72 K. The range of contractions was 2-13%, with the more polar cryosolutions contracting less. The potential uses of these results in the development of cryocooling conditions, as well as recent developments in determining minimum cryosolution soaking times, are discussed.

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“…Other methods of achieving satisfactory cryoprotection have been reported. It is possible, for example, to transfer some crystals from the normal mother liquor to unrelated solutions that contain cryoprotectant [14][15][16]. Another convenient approach is to grow the crystals in the presence of cryoprotectant, eliminating the need for a potentially damaging transfer to harvest buffer.…”

confidence: 99%

Cryocrystallography

1994

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Crystallographic studies are being extended to increasingly challenging systems: ever larger molecules, more complex viruses, membrane proteins, and assemblies. The crystals are often radiation-sensitive and scatter X-rays weakly, in many cases posing severe data collection problems. Synchrotron sources, high-performance area detectors and enhanced computational power have made it possible to tackle more difficult crystals and have greatly improved and simplified collection in general. Another tool, data collection at cryogenic temperatures, addresses more directly some of the problems associated with macromolecular crystals. Low-temperature techniques are coming into widespread use, and have become the routine method of data collection in a few laboratories. Cryogenic collection can make possible otherwise intractable projects, helping to expand the range of applications of crystallography to include some of the most exciting problems in biology. It is now clear that low-temperature techniques also offer real advantages with more conventional crystals, so that they should be considered for all macromolecular X-ray studies.

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Crystal transfer experiments carried out with crystals of tryanosomal triosephosphate isomerase (TIM)

Cited by 11 publications

References 18 publications

Structure-based approaches towards identification of fragments for the low-druggability ATAD2 bromodomain

Structure-based approaches towards identification of fragments for the low-druggability ATAD2 bromodomain

Progress in rational methods of cryoprotection in macromolecular crystallography

Cryocrystallography

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