Expression, purification, crystallization and structure determination of two glutathione S-transferase-like proteins from **Shewanella oneidensis**

Remmerie; Vandenbroucke, Karen; De Smet, Linda; Carpentier, Wesley; De Vos, Dirk; Stout, Jan; Van Beeumen, Jozef; Savvides, Savvas N.

doi:10.1107/S1744309108014589

Title

Expression, purification, crystallization and structure determination of two glutathione S-transferase-like proteins from **Shewanella oneidensis**

Author

Remmerie

Vandenbroucke, Karen

De Smet, Linda

Carpentier, Wesley

De Vos, Dirk

Stout, Jan

Van Beeumen, Jozef

Savvides, Savvas N.

Abstract

Genome analysis of Shewanella oneidensis, a Gram-negative bacterium with an unusual repertoire of respiratory and redox capabilities, revealed the presence of six glutathione S-transferase-like genes (sogst1sogst6). Glutathione S-transferases (GSTs; EC 2.5.1.18) are found in all kingdoms of life and are involved in phase II detoxification processes by catalyzing the nucleophilic attack of reduced glutathione on diverse electrophilic substrates, thereby decreasing their reactivity. Structurefunction studies of prokaryotic GST-like proteins are surprisingly underrepresented in the scientific literature when compared with eukaryotic GSTs. Here, the production and purification of recombinant SoGST3 (SO_1576) and SoGST6 (SO_4697), two of the six GST-like proteins in S. oneidensis, are reported and preliminary crystallographic studies of crystals of the recombinant enzymes are presented. SoGST3 was crystallized in two different crystal forms in the presence of GSH and DTT that diffracted to high resolution: a primitive trigonal form in space group P31 that exhibited merohedral twinning with a high twin fraction and a primitive monoclinic form in space group P21. SoGST6 yielded primitive orthorhombic crystals in space group P212121 from which diffraction data could be collected to medium resolution after application of cryo-annealing protocols. Crystal structures of both SoGST3 and SoGST6 have been determined based on marginal search models by maximum-likelihood molecular replacement as implemented in the program Phaser.

Language

English

Source (journal)

Acta crystallographica: section F: structural biology and crystallization communications. - Copenhagen

Publication

Copenhagen : 2008

ISSN

1744-3091

DOI

10.1107/S1744309108014589

Volume/pages

64 :6 (2008) , p. 548-553