Inducing phase changes in crystals of macromolecules: Status and perspectives for controlled crystal dehydration

Russi, Silvia; Juers, D.H.; Sánchez-Weatherby, Juan; Pellegrini, Erika; Mossou, Estelle; Forsyth, V. Trevor; Huet, Julien; Gobbo, Alexandre; Felisaz, Franck; Moya, Raphael; McSweeney, Seán; Cusack, Stephen; Cipriani, F.; Bowler, Matthew W.

doi:10.1016/j.jsb.2011.03.002

Cited by 53 publications

(60 citation statements)

References 56 publications

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“…4). All of these factors could have been facilitated by the presence of PEG in the crystallization medium, as has been suggested in other studies (Russi et al, 2011). The resilience of the hexagonal lattice to the changes in RH could be indicated by the relatively robust response to the increase in the number of waters per amino-acid residue (Fig.…”

Section: Discussionsupporting

confidence: 55%

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Elastic and inelastic diffraction changes upon variation of the relative humidity environment of PurE crystals

Oliete¹,

Pous

Rodríguez-Puente³

et al. 2013

Acta Crystallogr D Biol Crystallogr

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The different changes observed in the diffraction patterns of three different crystal forms (hexagonal, trigonal and monoclinic) of PurE (EC 4.1.1.21), an enzyme from the purine-biosynthesis pathway of Bacillus anthracis, upon a wide range of changes in the relative humidity environment of the crystals are documented. In addition, the changes in the unit-cell parameters, volume and bulk solvent in the three different crystal forms were systematically followed. In an attempt to explain the elastic (P6(5)22) and inelastic (P3(1)21) changes in the diffraction pattern, refined structures of the three different crystal forms determined at 100 K are presented, with particular emphasis on the tertiary and quaternary structural differences, crystal packing, intermolecular and intramolecular interactions and solvent structure. The refined structures show that the precipitant salts, solvent structure (both ordered and bulk) and conformation of the C-termini all play a role in creating a unique cement at both the intramolecular and intermolecular contacts of the different crystal forms. It is suggested that it is the combination of polyethylene glycol and the structure of the ordered water molecules (first and second layers) as well as the structure of the bulk solvent that are the critical factors in the plasticity of the hexagonal crystal packing as opposed to the inelastic responses of the lower symmetry forms.

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Section: Discussionsupporting

confidence: 55%

“…The current status of these developments in macromolecular crystallography have recently been reviewed (Russi et al, 2011). The importance of controlling the RH when soaking ligands into pre-grown apoenzyme crystals has been discussed in relation to fragment-based drug-discovery approaches (Bottcher et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

Elastic and inelastic diffraction changes upon variation of the relative humidity environment of PurE crystals

Oliete¹,

Pous

Rodríguez-Puente³

et al. 2013

Acta Crystallogr D Biol Crystallogr

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“…Mounted crystals were subjected to controlled dehydration 46 with a HC1c humidity control device at BESSY II beamline BL14.3 47 , reducing humidity from 93% to 89% in 1% steps with 2 min incubation prior to flash cooling in liquid N 2 . X-ray diffraction data were collected at 0.8726 Å wavelength at the ESRF ID23-2 microfocus beamline and processed in space group P2 using the XDS package 48 and TRUNCATE 49 yielding a final dataset at 2.1 Å resolution as judged from the Wilson plot (with Wilson B-factor 27.8), the CC 1/2 coefficient and model refinement statistics (see Table 1).…”

Section: Methodsmentioning

confidence: 99%

A mechanism for intracellular release of Na+ by neurotransmitter/sodium symporters

Malinauskaite

Quick

Reinhard

et al. 2014

Nat Struct Mol Biol

171

275

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Neurotransmitter:sodium symporters (NSS) terminate synaptic signal transmission by Na+-dependent reuptake of released neurotransmitters, with key conformational states reported for a bacterial homolog LeuT and an inhibitor-bound Drosophila dopamine transporter. However, a coherent mechanism of Na+-driven transport has not been described. Here, we present two crystal structures of MhsT, a NSS member from Bacillus halodurans, in occluded inward-facing states with bound Na+ ions and L-Trp that provide insight into the cytoplasmic release of Na+. The switch from outward- to inward-oriented states is centered on the partial unwinding of transmembrane helix 5, which is facilitated by a conserved GlyX9Pro motif that opens an intracellular pathway for water to access the Na2 site. Based on our structural and functional findings we propose a mechanism according to which solvation through the TM5 pathway facilitates Na+ release from Na2 and the transition to an inward-open state.

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“…Dehydration can increase the extent of crystal contacts and improve diffraction (Kiefersauer et al, 2000) (Russi et al, 2011), but can also be detrimental (Bernal and Crowfoot, 1934). The smaller cell volume may have been part of the reason tetragonal thermolysin required tert-butanol instead of methanol, and follow-up studies are being conducted to further understand this result.…”

Section: Resultsmentioning

confidence: 99%

Efficient cryoprotection of macromolecular crystals using vapor diffusion of volatile alcohols

Farley

Juers

2014

Journal of Structural Biology

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Macromolecular X-ray crystallography, usually done at cryogenic temperature to limit radiation damage, often requires liquid cryoprotective soaking that can be labor intensive and damaging to crystals. Here we describe a method for cryoprotection that uses vapor diffusion of volatile cryoprotective agents into loop-mounted crystals. The crystal is mounted into a vial containing a small volume of an alcohol-based cryosolution. After a short incubation with the looped crystal sitting in the cryosolution vapor, the crystal is transferred directly from the vial into the cooling medium. Effective for several different protein crystals, the approach obviates the need for liquid soaking and opens up a heretofore underutilized class of cryoprotective agents for macromolecular crystallography.

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Inducing phase changes in crystals of macromolecules: Status and perspectives for controlled crystal dehydration

Cited by 53 publications

References 56 publications

Elastic and inelastic diffraction changes upon variation of the relative humidity environment of PurE crystals

Elastic and inelastic diffraction changes upon variation of the relative humidity environment of PurE crystals

A mechanism for intracellular release of Na+ by neurotransmitter/sodium symporters

Efficient cryoprotection of macromolecular crystals using vapor diffusion of volatile alcohols

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