The Structure of an Orthorhombic Crystal Form of a 'forced Reduced' Thiol Peroxidase Reveals Lattice Formation Aided by the Presence of the Affinity Tag

Overview

Journal Acta Crystallogr Sect F Struct Biol Cryst Commun

Date 2012 Jun 14

PMID 22691780

Citations 2

Authors

Katherine S H Beckham

Olwyn Byron

Andrew J Roe

Mads Gabrielsen

Affiliations

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Abstract

Thiol peroxidase (Tpx) is an atypical 2-Cys peroxiredoxin, which has been suggested to be important for cell survival and virulence in Gram-negative pathogens. The structure of a catalytically inactive version of this protein in an orthorhombic crystal form has been determined by molecular replacement. Structural alignments revealed that Tpx is conserved. Analysis of the crystal packing shows that the linker region of the affinity tag is important for formation of the crystal lattice.

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References

Horst S, Jaeger T, Denkel L, Rouf S, Rhen M, Bange F . Thiol peroxidase protects Salmonella enterica from hydrogen peroxide stress in vitro and facilitates intracellular growth. J Bacteriol. 2010; 192(11):2929-32. PMC: 2876503. DOI: 10.1128/JB.01652-09. View

Baron C . Antivirulence drugs to target bacterial secretion systems. Curr Opin Microbiol. 2010; 13(1):100-5. DOI: 10.1016/j.mib.2009.12.003. View

Krissinel E, Henrick K . Inference of macromolecular assemblies from crystalline state. J Mol Biol. 2007; 372(3):774-97. DOI: 10.1016/j.jmb.2007.05.022. View

Velankar S, Alhroub Y, Best C, Caboche S, Conroy M, Dana J . PDBe: Protein Data Bank in Europe. Nucleic Acids Res. 2011; 40(Database issue):D445-52. PMC: 3245096. DOI: 10.1093/nar/gkr998. View

Emsley P, Lohkamp B, Scott W, Cowtan K . Features and development of Coot. Acta Crystallogr D Biol Crystallogr. 2010; 66(Pt 4):486-501. PMC: 2852313. DOI: 10.1107/S0907444910007493. View

Wang D, Zetterstrom C, Gabrielsen M, Beckham K, Tree J, Macdonald S . Identification of bacterial target proteins for the salicylidene acylhydrazide class of virulence-blocking compounds. J Biol Chem. 2011; 286(34):29922-31. PMC: 3191033. DOI: 10.1074/jbc.M111.233858. View

Cha M, Kim H, Kim I . Thioredoxin-linked "thiol peroxidase" from periplasmic space of Escherichia coli. J Biol Chem. 1995; 270(48):28635-41. DOI: 10.1074/jbc.270.48.28635. View

Murshudov G, Skubak P, Lebedev A, Pannu N, Steiner R, Nicholls R . REFMAC5 for the refinement of macromolecular crystal structures. Acta Crystallogr D Biol Crystallogr. 2011; 67(Pt 4):355-67. PMC: 3069751. DOI: 10.1107/S0907444911001314. View

Evans P . Scaling and assessment of data quality. Acta Crystallogr D Biol Crystallogr. 2005; 62(Pt 1):72-82. DOI: 10.1107/S0907444905036693. View

10.

Chen V, Arendall 3rd W, Headd J, Keedy D, Immormino R, Kapral G . MolProbity: all-atom structure validation for macromolecular crystallography. Acta Crystallogr D Biol Crystallogr. 2010; 66(Pt 1):12-21. PMC: 2803126. DOI: 10.1107/S0907444909042073. View

11.

Baker L, Poole L . Catalytic mechanism of thiol peroxidase from Escherichia coli. Sulfenic acid formation and overoxidation of essential CYS61. J Biol Chem. 2003; 278(11):9203-11. PMC: 3845838. DOI: 10.1074/jbc.M209888200. View

12.

Gabrielsen M, Zetterstrom C, Wang D, Beckham K, Elofsson M, Isaacs N . Expression, purification, crystallization and initial X-ray diffraction analysis of thiol peroxidase from Yersinia pseudotuberculosis. Acta Crystallogr Sect F Struct Biol Cryst Commun. 2010; 66(Pt 12):1606-9. PMC: 2998365. DOI: 10.1107/S1744309110039679. View

13.

Gabrielsen M, Beckham K, Feher V, Zetterstrom C, Wang D, Muller S . Structural characterisation of Tpx from Yersinia pseudotuberculosis reveals insights into the binding of salicylidene acylhydrazide compounds. PLoS One. 2012; 7(2):e32217. PMC: 3288085. DOI: 10.1371/journal.pone.0032217. View

14.

Bond C, Schuttelkopf A . ALINE: a WYSIWYG protein-sequence alignment editor for publication-quality alignments. Acta Crystallogr D Biol Crystallogr. 2009; 65(Pt 5):510-2. DOI: 10.1107/S0907444909007835. View

15.

Hall A, Sankaran B, Poole L, Karplus P . Structural changes common to catalysis in the Tpx peroxiredoxin subfamily. J Mol Biol. 2009; 393(4):867-81. PMC: 3664093. DOI: 10.1016/j.jmb.2009.08.040. View