Cycling Temperature Strategy: A Method to Improve the Efficiency of Crystallization Condition Screening of Proteins

Zhang, Chenyan; Yin, Da‐Chuan; Lu, Qi; Guo, Yang; Guo, Weihong; Wang, Xi-Kai; Li, Haisheng; Lu, Hui‐Meng; Ye, Ya-Jing

doi:10.1021/cg800689j

Cited by 37 publications

(33 citation statements)

References 32 publications

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“…The scan areas are (a) and (b) 23 Â 23 nm, (c) 20 Â 20 nm and (d) 34 Â Manipulation of temperature by cycling of a crystallization matrix is another way to utilize temperature to induce nucleation. With this approach, the crystallization experiment is initially prepared at 4 C in an incubator that is subsequently temperature ramped to 30 C over a period of 24 h and then ramped back to 4 C over another 24 h. During the temperature cycling the experiment is scored at 4, 20 and 30 C and again at 4 C. Variations can be introduced as shorter or longer ramping periods between temperature points, as well as evaluating different hold times at desired temperatures (Zhang et al, 2008).…”

Section: Optimization Of Temperaturementioning

confidence: 99%

Optimization of crystallization conditions for biological macromolecules

McPherson

Cudney²

2014

Acta Crystallogr F Struct Biol Commun

106

112

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For the successful X-ray structure determination of macromolecules, it is first necessary to identify, usually by matrix screening, conditions that yield some sort of crystals. Initial crystals are frequently microcrystals or clusters, and often have unfavorable morphologies or yield poor diffraction intensities. It is therefore generally necessary to improve upon these initial conditions in order to obtain better crystals of sufficient quality for X-ray data collection. Even when the initial samples are suitable, often marginally, refinement of conditions is recommended in order to obtain the highest quality crystals that can be grown. The quality of an X-ray structure determination is directly correlated with the size and the perfection of the crystalline samples; thus, refinement of conditions should always be a primary component of crystal growth. The improvement process is referred to as optimization, and it entails sequential, incremental changes in the chemical parameters that influence crystallization, such as pH, ionic strength and precipitant concentration, as well as physical parameters such as temperature, sample volume and overall methodology. It also includes the application of some unique procedures and approaches, and the addition of novel components such as detergents, ligands or other small molecules that may enhance nucleation or crystal development. Here, an attempt is made to provide guidance on how optimization might best be applied to crystal-growth problems, and what parameters and factors might most profitably be explored to accelerate and achieve success.

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Section: Optimization Of Temperaturementioning

confidence: 99%

Optimization of crystallization conditions for biological macromolecules

McPherson

Cudney²

2014

Acta Crystallogr F Struct Biol Commun

106

112

View full text Add to dashboard Cite

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“…The crystallization plates (40-well plates with a diameter of 7 mm and depth of 10 mm) were purchased from Keyu Company, Jiangsu, China. To ensure accurate temperature control, the crystallization plates were placed in a homemade chamber (150 × 150 × 150 mm 3 ) wrapped with a water-jacket in which temperature-controlled water circulated from a water bath (PolyScience 9712, PolyScience, USA), which controlled the water temperature within ±0.1 K. A more detailed description of the temperature control apparatus can be found in [16]. Four days after the beginning of the crystallization experiments, the crystallization plates were removed from © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.crt-journal.org the temperature-controlled chamber and checked by a stereomicroscope (Nikon SMZ1000, Nikon, Japan).…”

Section: Methodsmentioning

confidence: 99%

Enhancement of nucleation during hanging drop protein crystallization using HF Treatment of cover glasses

Guo¹,

Yin²,

Lu³

et al. 2010

Cryst. Res. Technol.

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We examined a simple approach, i.e., etching cover glasses using hydrofluoric acid (HF), to determine whether cover glass treatment enhances nucleation in hanging drop protein crystallization. Hen egg white lysozyme and proteinase K were used as the model proteins. We found that the treatment increased the success rate of crystallization. The results indicated that the simple treatment, which is easy to adopt without changing much in the hanging drop method, can be utilized as an alternative method to enhance protein crystallization screens.

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“…At a specific protein concentration, the lower the solubility is, the higher the supersaturation will be (Chen et al, 2012;Yin et al, 2002;Zhang et al, 2008). Therefore, it was previously believed that the optimal pH for protein crystallization was the pI value of the protein owing to the pronounced solubility minimum (Rupp, 2009).…”

Section: If Only One Protein Solution Ph Is Allowedmentioning

confidence: 99%

A strategy for selecting the pH of protein solutions to enhance crystallization

Zhang

Yin

et al. 2013

Acta Cryst Sect F

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The pH of a solution is an important parameter in crystallization that needs to be controlled in order to ensure success. The actual pH of the crystallization droplet is determined by the combined contribution of the buffers in the screening and protein solutions, although the contribution of the latter to the pH is often ignored. In this study, the effects of the buffer and protein solution pH values on the results of screening are systematically investigated. It was found that these parameters significantly affected the results and thus the following strategy for the selection of appropriate pH values is proposed: (i) when screening with only one protein solution, the pH should be as low, as high or as divergent from the pI as possible for a basic, acidic or neutral protein, respectively, within its stable pH range; (ii) when screening with two protein solutions, the pH values should be well separated from one another; and (iii) when multiple pH values are utilized, an even distribution of pH values is the best approach to increase the success rate of crystallization.

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Cycling Temperature Strategy: A Method to Improve the Efficiency of Crystallization Condition Screening of Proteins

Cited by 37 publications

References 32 publications

Optimization of crystallization conditions for biological macromolecules

Optimization of crystallization conditions for biological macromolecules

Enhancement of nucleation during hanging drop protein crystallization using HF Treatment of cover glasses

A strategy for selecting the pH of protein solutions to enhance crystallization

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