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Int. J. Mol. Sci. 2011, 12(9), 6312-6319; doi:10.3390/ijms12096312
Communication

Improving Protein Crystal Quality by the Without-Oil Microbatch Method: Crystallization and Preliminary X-ray Diffraction Analysis of Glutathione Synthetase from Pseudoalteromonas haloplanktis

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Received: 26 July 2011; in revised form: 5 September 2011 / Accepted: 14 September 2011 / Published: 23 September 2011
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
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Abstract: Glutathione synthetases catalyze the ATP-dependent synthesis of glutathione from L-γ-glutamyl-L-cysteine and glycine. Although these enzymes have been sequenced and characterized from a variety of biological sources, their exact catalytic mechanism is not fully understood and nothing is known about their adaptation at extremophilic environments. Glutathione synthetase from the Antarctic eubacterium Pseudoalteromonas haloplanktis (PhGshB) has been expressed, purified and successfully crystallized. An overall improvement of the crystal quality has been obtained by adapting the crystal growth conditions found with vapor diffusion experiments to the without-oil microbatch method. The best crystals of PhGshB diffract to 2.34 Å resolution and belong to space group P212121, with unit-cell parameters a = 83.28 Å, b = 119.88 Å, c = 159.82 Å. Refinement of the model, obtained using phases derived from the structure of the same enzyme from Escherichia coli by molecular replacement, is in progress. The structural determination will provide the first structural characterization of a psychrophilic glutathione synthetase reported to date.
Keywords: crystal quality; without-oil microbatch; glutathione synthetase; psychrophile; X-ray crystallography crystal quality; without-oil microbatch; glutathione synthetase; psychrophile; X-ray crystallography
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Merlino, A.; Krauss, I.R.; Albino, A.; Pica, A.; Vergara, A.; Masullo, M.; Vendittis, E.D.; Sica, F. Improving Protein Crystal Quality by the Without-Oil Microbatch Method: Crystallization and Preliminary X-ray Diffraction Analysis of Glutathione Synthetase from Pseudoalteromonas haloplanktis. Int. J. Mol. Sci. 2011, 12, 6312-6319.

AMA Style

Merlino A, Krauss IR, Albino A, Pica A, Vergara A, Masullo M, Vendittis ED, Sica F. Improving Protein Crystal Quality by the Without-Oil Microbatch Method: Crystallization and Preliminary X-ray Diffraction Analysis of Glutathione Synthetase from Pseudoalteromonas haloplanktis. International Journal of Molecular Sciences. 2011; 12(9):6312-6319.

Chicago/Turabian Style

Merlino, Antonello; Krauss, Irene Russo; Albino, Antonella; Pica, Andrea; Vergara, Alessandro; Masullo, Mariorosario; Vendittis, Emmanuele De; Sica, Filomena. 2011. "Improving Protein Crystal Quality by the Without-Oil Microbatch Method: Crystallization and Preliminary X-ray Diffraction Analysis of Glutathione Synthetase from Pseudoalteromonas haloplanktis." Int. J. Mol. Sci. 12, no. 9: 6312-6319.



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