Purification, crystallization and preliminary X-ray analysis of native and selenomethionine class I tagatose-1,6-bisphosphate aldolase from Streptococcus pyogenes
| Autor: | Brigitte Liotard, Jurgen Sygusch |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Streptococcus pyogenes
Recombinant Fusion Proteins medicine.disease_cause Cleavage (embryo) Crystallography X-Ray chemistry.chemical_compound Bacterial Proteins Structural Biology Glyceraldehyde medicine Escherichia coli Selenomethionine Dihydroxyacetone phosphate Aldehyde-Lyases Hexoses biology Hydrophilic interaction chromatography Aldolase A General Medicine chemistry Biochemistry biology.protein Crystallization Tagatose |
| Zdroj: | Acta crystallographica. Section D, Biological crystallography. 60(Pt 3) |
| ISSN: | 0907-4449 |
| Popis: | 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 P2(1)2(1)2(1), with unit-cell parameters a = 63.7, b = 108.1, c = 238.7 A for the native form and a = 64.1, b = 108.3, c = 239.8 A for the selenomethionine derivative. The asymmetric unit contains four protomers, corresponding to a crystal volume per protein weight (V(M)) of 2.8 A(3) Da(-1) and a solvent content of 56% by volume. |
| Databáze: | OpenAIRE |
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