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

1 Dipartimento di Chimica “Paolo Corradini”, Università di Napoli Federico II, Complesso Universitario di Monte Sant’Angelo, Via Cinthia, Naples I-80126, Italy 2 Istituto di Biostrutture e Bioimmagini, CNR, Via Mezzocannone 16, Naples I-80134, Italy 3 Dipartimento di Biochimica e Biotecnologie Mediche, Università di Napoli Federico II, Via Pansini 5, Naples I-80131, Italy 4 Dipartimento di Studi delle Istituzioni e dei Sistemi Territoriali, Università di Napoli “Parthenope”, Via Medina 40, Naples I-80133, Italy
* Author to whom correspondence should be addressed.
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)
PDF Full-text Download PDF Full-Text [253 KB, uploaded 23 September 2011 10:59 CEST]
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

Article Statistics

Load and display the download statistics.

Citations to this Article

Cite This Article

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.

Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert