Match!

A New Design of Protein Crystallization Plates To Expand Concentration Screening Space in Cross-Diffusion Microbatch and Microbatch Methods

Published on Feb 3, 2016in Crystal Growth & Design4.153
· DOI :10.1021/acs.cgd.5b00702
Chen Dong4
Estimated H-index: 4
,
Chen-Yan Zhang9
Estimated H-index: 9
+ 3 AuthorsDa-Chuan Yin18
Estimated H-index: 18
Abstract
Protein crystallizes at specific conditions (correct precipitants, appropriate concentrations of protein and precipitants, suitable pH and correct temperature, etc.). If the conditions are not appropriate, crystallization will not occur. In protein crystallization screening, the target protein is mixed one by one with many chemical agents and then incubated at a set temperature. If the concentrations of the chemical agents and the target proteins are not in a range suitable for crystallization, the crystallization will not occur. To expand the concentration screening space, we propose in this paper a new design of protein crystallization plates for cross-diffusion microbatch and microbatch methods. The new plates have 96 units corresponding to the conditions of the commercial screening kits, and each unit contains four wells for holding the crystallization droplets. By dispensing crystallization droplets to the four wells at different volume ratios of protein to precipitant solutions, we can obtain four d...
  • References (34)
  • Citations (4)
📖 Papers frequently viewed together
56 Citations
6 Citations
1 Citations
78% of Scinapse members use related papers. After signing in, all features are FREE.
References34
Newest
#1Sahir KhurshidH-Index: 9
Last. Naomi E. ChayenH-Index: 33
view all 4 authors...
A Protocol for obtaining high-quality protein crystals that involves addition to the protein solution of solid or semi-liquid nucleants that provide the experimenter the means to control crystal growth.
52 CitationsSource
#1Rui-Qing Chen (NPU: Northwestern Polytechnical University)H-Index: 5
#2Da-Chuan Yin (NPU: Northwestern Polytechnical University)H-Index: 18
Last. Yue Liu (NPU: Northwestern Polytechnical University)H-Index: 4
view all 6 authors...
6 CitationsSource
#1Qin-Qin Lu (NPU: Northwestern Polytechnical University)H-Index: 8
#2Xu-Zhuo Xie (NPU: Northwestern Polytechnical University)H-Index: 3
Last. Da-Chuan Yin (NPU: Northwestern Polytechnical University)H-Index: 18
view all 7 authors...
The gradual desiccation method (GDM) is a modification of the vapor diffusion method for protein crystallization screening. This method can dramatically increase the chances of obtaining protein crystals and is therefore potentially useful for practical protein crystallization screening. However, it is troublesome to prepare the desiccant for the GDM because each of the 96 desiccants must be of the same mass. Repeated manual weighing of the desiccant (at least 96 times for one plate) to obtain t...
1 CitationsSource
#1Chen-Yan ZhangH-Index: 2
#2Mausumi MazumdarH-Index: 6
Last. Sheng-Xiang LinH-Index: 24
view all 5 authors...
The nucleation zone has to be reached for any crystal to grow, and the search for crystallization conditions of new proteins is a trial and error process. Here a convenient screening strategy is studied in detail that varies the volume ratio of protein sample to the reservoir solution in the drop to initiate crystallization that is named “composition modification”. It is applied after the first screen and has been studied with twelve proteins. Statistical analysis shows a significant improvement...
1 CitationsSource
#1Janet Newman (CSIRO: Commonwealth Scientific and Industrial Research Organisation)H-Index: 27
Abstract Turning commercial lab automation into a high-throughput centre requires an underlying process, and implementing checks to ensure that the process is working as it should. At the Collaborative Crystallisation Centre (C3), protein samples from local, national and international groups are set up in crystallisation screening and optimisation experiments with two thousand 96 well plates being set up each year. During its five years of operation, the C3 has implemented a series of enabling p...
23 CitationsSource
#1Qin-Qin Lu (NPU: Northwestern Polytechnical University)H-Index: 8
#2Da-Chuan Yin (NPU: Northwestern Polytechnical University)H-Index: 18
Last. Rui-Qing Chen (NPU: Northwestern Polytechnical University)H-Index: 5
view all 5 authors...
In this study, effects of diluting either protein or crystallization agents in the droplets on the success rate of protein crystallization was investigated. Diluting the crystallization agent was found to increase the success rate of protein crystallization. Theoretical analysis showed that, concentration ranges of both protein and crystallization agent that can be scanned during the vapor diffusion process are wider with diluting the crystallization agent than that without dilution, resulting i...
3 CitationsSource
#1Liang LiH-Index: 65
#2Rustem F. Ismagilov (U of C: University of Chicago)H-Index: 63
To obtain protein crystals, researchers must search for conditions in multidimensional chemical space. Empirically, thousands of crystallization experiments are carried out to screen various precipitants at multiple concentrations. Microfluidics can manipulate fluids on a nanoliter scale, and it affects crystallization twofold. First, it miniaturizes the experiments that can currently be done on a larger scale and enables crystallization of proteins that are available only in small amounts. Seco...
121 CitationsSource
#1F. Gorrec (LMB: Laboratory of Molecular Biology)H-Index: 8
A 96-condition initial screen for protein crystallization, called MORPHEUS, has been developed at the MRC Laboratory of Molecular Biology, Cambridge, England (MRC-LMB). The concept integrates several innovative approaches, such as chemically compatible mixes of potential ligands, new buffer systems and precipitant mixes that also act as cryoprotectants. Instead of gathering a set of crystallization conditions that have already been successful, a selection of molecules frequently observed in the ...
135 CitationsSource
#1Fermín Otálora (UGR: University of Granada)H-Index: 23
#2Jose A. Gavira (UGR: University of Granada)H-Index: 24
Last. Juan Manuel García-Ruiz (UGR: University of Granada)H-Index: 31
view all 4 authors...
Accumulated experience during the last years on counterdiffusion crystallization methods shows that they are a convenient and generally applicable way of optimizing solution crystal growth experiments. Irrespective of whether the objective of the experiment is to improve crystal quality or size, many experiments reporting a positive or neutral effect of counterdiffusion exists, but adverse effects are consistently absent. Thus counterdiffusion is viewed as a rational crystallization approach to ...
83 CitationsSource
#1Renaud Vincentelli (CNRS: Centre national de la recherche scientifique)H-Index: 24
#2Stéphane Canaan (CNRS: Centre national de la recherche scientifique)H-Index: 24
Last. Christophe Bignon (CNRS: Centre national de la recherche scientifique)H-Index: 14
view all 10 authors...
One of the main stumbling blocks encountered when attempting to express foreign proteins in Escherichia coli is the occurrence of amorphous aggregates of misfolded proteins, called inclusion bodies (IB). Developing efficient protein native structure recovery procedures based on IB refolding is therefore an important challenge. Unfortunately, there is no “universal” refolding buffer: Experience shows that refolding buffer composition varies from one protein to another. In addition, the methods de...
137 CitationsSource
Cited By4
Newest
#1Hai Hou (NPU: Northwestern Polytechnical University)H-Index: 1
#2Miao Shi (NPU: Northwestern Polytechnical University)H-Index: 1
Last. Da-Chuan Yin (NPU: Northwestern Polytechnical University)H-Index: 18
view all 11 authors...
The wells of the novel plate are pre-embedded with cross-linked protein microcrystals which can act as the seeds for protein crystallization. The results of crystallization screening experiments showed that the novel plate exhibited dramatically improved performance in finding crystallization conditions as compared with the original CDM plate.
1 CitationsSource
#1Xue-Zhou YangH-Index: 3
#2Chen-Yan ZhangH-Index: 9
Last. Da-Chuan YinH-Index: 18
view all 9 authors...
Finding new nucleants to promote protein crystallization is an important task, especially for purposes other than structural determination. Here, we investigated cyclodextrins and its derivative particles, as potential nucleants for protein crystallization. β-cyclodextrin (β-CD) and its derivatives (including p-toluenesulfonyl-β-cyclodextrin (PTCD), polymer-β-cyclodextrin (PCD), Mono-(6-(1,6-hexamethylenediamine)-6-deoxy)-β-Cyclodextrin (MHCD) and Mercapto-β-cyclodextrin (MCD)) were used as nucl...
4 CitationsSource
#1Chen-Yan Zhang (NPU: Northwestern Polytechnical University)H-Index: 9
#2Chen Dong (NPU: Northwestern Polytechnical University)H-Index: 4
Last. Da-Chuan Yin (NPU: Northwestern Polytechnical University)H-Index: 18
view all 9 authors...
Abstract We have proposed a rational strategy for selecting a suitable pH of protein solution based on protein biochemical properties. However, it is difficult to use this strategy for biochemical properties unknown proteins. In this paper, a simpler and faster pH buffer strategy was proposed. An additional pH-controlling buffer was added to crystallization droplet mixed with protein solution and commercial crystallization reagents to adjust its pH. The results revealed that protein crystallizat...
1 CitationsSource
#1Rui-Qing Chen (NPU: Northwestern Polytechnical University)H-Index: 5
#2Qing-Di Cheng (NPU: Northwestern Polytechnical University)H-Index: 4
Last. Da-Chuan Yin (NPU: Northwestern Polytechnical University)H-Index: 18
view all 8 authors...
Protein crystallization occurs when the solution conditions are suitable for nucleation and growth of the crystals. pH, as one of the most important parameters that governs the protein crystallization process, can affect the conformation, activity, electrostatic interactions and solubility of protein in the solution. Hence, manipulating the pH is an important path to success in crystallizing proteins. In this paper, we show that varying pH levels over the incubation time during the crystallizati...
3 CitationsSource