Soaking of crystals for macromolecular crystallography in a capillary

Lebioda, Lukasz; Zhang, E.

doi:10.1107/s0021889891011342

Cited by 12 publications

(13 citation statements)

References 1 publication

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“…To complete the Na ϩ -K ϩ exchange, the crystals were transferred directly into 1 ml of the potassium solution. The Na ϩ -Rb ϩ exchange was accomplished using a capillary soaking method (18) with the potassium phosphate solution containing RbCl instead of KCl. The crystal was placed in a capillary tube used to mount crystals for diffraction purposes.…”

Section: Methodsmentioning

confidence: 99%

The Na+ Binding Site of Thrombin

Guinto

Vindigni

et al. 1995

Journal of Biological Chemistry

241

258

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Thrombin is an allosteric serine protease existing in two forms, slow and fast, targeted toward anticoagulant and procoagulant activities. The slow 3 fast transition is induced by Na , and Lys 224 , and by three highly conserved water molecules in the D-Phe-Pro-Arg chloromethylketone thrombin. The sequence in the Na ؉ binding loop is highly conserved in thrombin from 11 different species and is homologous to that found in other serine proteases involved in blood coagulation. Mutation of two Asp residues flanking Arg 221a (D221A/D222K) almost abolishes the allosteric properties of thrombin and shows that the Na ؉ binding loop is also involved in direct recognition of protein C and antithrombin.

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Section: Methodsmentioning

confidence: 99%

The Na+ Binding Site of Thrombin

Guinto

Vindigni

et al. 1995

Journal of Biological Chemistry

241

258

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“…The same capillary used as a growth cell can be used later for soaking crystals with ligands prior to X-ray studies because, as demonstrated by Lebiola & Zhang (1992), equilibration of the ligand is correctly attained by diffusion through the mother liquid into a capillary volume. It also seems clear that this technique can be used successfully for the screening of crystallization conditions and for in situ diffusion of inhibitors, heavy atoms and substrates.…”

Section: Discussionmentioning

confidence: 99%

Investigations on protein crystal growth by the gel acupuncture method

García‐Ruiz

Moreno²

1994

Acta Crystallogr D Biol Crystallogr

118

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In this work we explore the possibilities of the gelacupuncture technique, proposed previously for the growth of protein single crystals [Garcia-Ruiz, Moreno, Viedma & Coll (1993). Mater. Res. Bull. 28,[541][542][543][544][545][546]. The main advantage of the technique is that the crystals are obtained inside an X-ray capillary and, unlike classical microdiffusion techniques, it involves a very simple and accurate technical arrangement that permits the continuous monitoring of the crystals in their growth environment. In particular, we describe the growth of single crystals of lysozyme, concanavalin A and ribonuclease A. Different starting conditions have been used to grow single crystals of these proteins into different types of capillaries at several protein and precipitating-agent concentrations. It is demonstrated that the technique works for a wide range of precipitating agents commonly used in protein crystal growth, such as large polymers (PEG 4000 and PEG 6000), organic solvents (from methanol to butanol) and salts [NaC1, (NH4)2SO4]. The range of inner diameter of the capillaries for which the technique works correctly has been also studied. The growth process and possible crystal movement was followed by video microscopy. Lysozyme crystals up to 3.1 mm were obtained but the average maximum linear crystal sizes were 2.0 mm for lysozyme, 0.4 mm for concanavalin A and 1.2 mm for ribonuclease, respectively. The waiting times to reach such a size, measured from the set-up of the experiments, were 72 h, 10 d and 5 d, respectively. Gels of tetramethoxysilane, tetraethoxysilane, sodium silicate, agar, high-strength agar and gel-gro have been tested in relation to their mechanical properties and their chemical interaction with the reactants. Finally, we discuss briefly the advantages of the gel-acupuncture technique and plausible applications other than crystal growth.

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“…The potential to carry out halide exchange processes within this family of complexes was explored by soaking crystals 38 (Table 6). Increasing the soaking period to 11 days led to a larger increase in the mole fraction of iodide in the complex from 0.39 to 0.70 ( Table 6).…”

Section: Halide Exchange Processesmentioning

confidence: 99%

Complexes of Thiourea with Alkali Metal Bromides and Iodides: Structural Properties, Mixed-Halide and Mixed-Metal Materials, and Halide Exchange Processes

Charalampopoulos

Kariuki

Harris

2017

Crystal Growth & Design

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We report the preparation and structural properties of complexes of metal halides and thiourea with composition MX[thiourea]4 (MX = KBr, KI, RbI, CsI), together with the mixed-halide materials KBrnI1-n[thiourea]4 (0 < n < 1) and the mixed-metal materials KnCs1-nI[thiourea]4 (0 < n < 1). These materials are isostructural, with a tetragonal structure (space group P4/mnc) characterized by M + [thiourea]4 coordination columns along the 4-fold axis and halide anions located in channels in the region of space between adjacent columns and running parallel to the columns. For the mixed-halide materials, the stoichiometry KBrnI1-n[thiourea]4 depends on the bromide/iodide ratio in the crystallization solution; the crystalline complexes have a higher bromide/iodide ratio than the crystallization solution, indicating preferential incorporation of bromide within the complex. Soaking crystals of KBr0.61I0.39[thiourea]4 in iodide containing solutions leads to halide exchange, with the iodide to bromide ratio increasing relative to the parent crystal. Further experiments produced no evidence that these thiourea complexes can accommodate extended polyiodide networks.

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Soaking of crystals for macromolecular crystallography in a capillary

Cited by 12 publications

References 1 publication

The Na+ Binding Site of Thrombin

The Na+ Binding Site of Thrombin

Investigations on protein crystal growth by the gel acupuncture method

Complexes of Thiourea with Alkali Metal Bromides and Iodides: Structural Properties, Mixed-Halide and Mixed-Metal Materials, and Halide Exchange Processes

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