IngentaConnect Purification, crystallization and preliminary X-ray analysis of n...

Authors: Liotard, Brigitte; Sygusch, Jurgen

Source: Acta Crystallographica Section D, Volume 60, Number 3, March 2004 , pp. 528-530(3)

Abstract:

Tagatose-1,6-bisphosphate aldolase (EC 4.1.2.40) is situated at the branching of the tagatose-6-phosphate and Embden-Meyerhof-Parnas (glycolysis) metabolic pathways, where it catalyzes the reversible cleavage of tagatose-1,6-bisphosphate to dihydroxyacetone phosphate and glyceraldehyde 3-phosphate. The recombinant protein from Streptococcus pyogenes was overexpressed in Escherichia coli in its native and selenomethionine-derivative forms and purified using ion-exchange and hydrophobic interaction chromatography. Orthorhombic crystals suitable for structural analysis were obtained by the hanging-drop vapour-diffusion method for both isoforms. The crystals belong to space group P2₁2₁2₁, with unit-cell parameters a = 63.7, b = 108.1, c = 238.7 Å for the native form and a = 64.1, b = 108.3, c = 239.8 Å for the selenomethionine derivative. The asymmetric unit contains four protomers, corresponding to a crystal volume per protein weight (V_M) of 2.8 Å³ Da⁻¹ and a solvent content of 56% by volume.

Keywords: tagatose-1,6-bisphosphate aldolase

Document Type: Research article

DOI: 10.1107/S0907444903028427

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Purification, crystallization and preliminary X-ray analysis of native and selenomethionine class I tagatose-1,6-bisphosphate aldolase from Streptococcus pyogenes