Cryogenic x-ray diffraction microscopy utilizing high-pressure cryopreservation

Lima, Enju; Chushkin, Yuriy; Linden, P. van der; Kim, Chae Un; Zontone, Federico; Carpentier, Philippe; Grüner, Sol M.; Pernot, Pétra

doi:10.1103/physreve.90.042713

Cited by 20 publications

(5 citation statements)

References 35 publications

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“…Previously reported benefits of high-pressure cryocooling include reduction of damage due to cryocooling (Kim et al, 2005), ability to cryocool samples other than crystals mounted in loops (Kim et al, 2007;Lima et al, 2014), stabilization of ligands (Albright et al, 2006), trapping of gas molecules [noble gases for phasing (Kim et al, 2006) as well as reactants in active sites (Domsic et al, 2008)] and observation of pressure effects on molecular structure (Barstow et al, 2009). We now add the use of HPC to reduce crystal disorder.…”

Section: Discussionmentioning

confidence: 99%

Reduction of lattice disorder in protein crystals by high-pressure cryocooling

et al. 2016

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High-pressure cryocooling (HPC) has been developed as a technique for reducing the damage that frequently occurs when macromolecular crystals are cryocooled at ambient pressure. Crystals are typically pressurized at around 200 MPa and then cooled to liquid nitrogen temperature under pressure; this process reduces the need for penetrating cryoprotectants, as well as the damage due to cryocooling, but does not improve the diffraction quality of the as-grown crystals. Here it is reported that HPC using a pressure above 300 MPa can reduce lattice disorder, in the form of high mosaicity and/or nonmerohedral twinning, in crystals of three different proteins, namely human glutaminase C, the GTP pyrophosphokinase YjbM and the uncharacterized protein lpg1496. Pressure lower than 250 MPa does not induce this transformation, even with a prolonged pressurization time. These results indicate that HPC at elevated pressures can be a useful tool for improving crystal packing and hence the quality of the diffraction data collected from pressurized crystals.

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Section: Discussionmentioning

confidence: 99%

Reduction of lattice disorder in protein crystals by high-pressure cryocooling

et al. 2016

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“…In comparison with electrons, X-rays are able to provide higher penetration, so that the sample does not have to be sectioned. Cryo-imaging has already been shown to be effective for biological samples using soft X-ray tomography (Schneider et al, 2010), hard X-ray CDI (Giewekemeyer et al, 2015;Huang et al, 2009;Lima et al, 2009Lima et al, , 2014Rodriguez et al, 2015) and ptychographic methods (Deng et al, 2015;Lima et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

Procedures for cryogenic X-ray ptychographic imaging of biological samples

Yusuf

Zhang

Chen

et al. 2017

Int Union Crystallogr J

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“…the limited pressure attainable and the escape of the gas from crystals during the pressure-release operation prior to flash cooling). The technical design of the cell is partly based on those of cryoprotectant-free highpressure cooling benches for biological crystals that have recently been developed (Kim et al, 2006(Kim et al, , 2005Lima et al, 2014; and, crucially, is compatible with SPINE standard sample supports (Cipriani et al, 2006), thus allowing the use of robotic sample changers during experimental sessions. However, the thermodynamic principle Table 1 Thermodynamic properties of selected gases.…”

Section: Introductionmentioning

confidence: 99%

Gas-sensitive biological crystals processed in pressurized oxygen and krypton atmospheres: deciphering gas channels in proteins using a novel `soak-and-freeze' methodology

Lafumat¹,

Mueller-Dieckmann²,

Leonard³

et al. 2016

J Appl Crystallogr

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International audienceMolecular oxygen (O2) is a key player in many fundamental biological processes. However, the combination of the labile nature and poor affinity of O2 often makes this substrate difficult to introduce into crystals at sufficient concentrations to enable protein/O2 interactions to be deciphered in sufficient detail. To overcome this problem, a gas pressure cell has been developed specifically for the `soak-and-freeze' preparation of crystals of O2-dependent biological molecules. The `soak-and-freeze' method uses high pressure to introduce oxygen molecules or krypton atoms (O2 mimics) into crystals which, still under high pressure, are then cryocooled for X-ray data collection. Here, a proof of principle of the gas pressure cell and the methodology developed is demonstrated with crystals of enzymes (lysozyme, thermolysin and urate oxidase) that are known to absorb and bind molecular oxygen and/or krypton. The successful results of these experiments lead to the suggestion that the soak-and-freeze method could be extended to studies involving a wide range of gases of biological, medical and/or environmental interest, including carbon monoxide, ethylene, methane and many others

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Cryogenic x-ray diffraction microscopy utilizing high-pressure cryopreservation

Cited by 20 publications

References 35 publications

Reduction of lattice disorder in protein crystals by high-pressure cryocooling

Reduction of lattice disorder in protein crystals by high-pressure cryocooling

Procedures for cryogenic X-ray ptychographic imaging of biological samples

Gas-sensitive biological crystals processed in pressurized oxygen and krypton atmospheres: deciphering gas channels in proteins using a novel `soak-and-freeze' methodology

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