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A complex iron-calcium cofactor catalyzing phosphotransfer chemistry.

Science (New York, N.Y.) (2014-09-06)
Shee Chien Yong, Pietro Roversi, James Lillington, Fernanda Rodriguez, Martin Krehenbrink, Oliver B Zeldin, Elspeth F Garman, Susan M Lea, Ben C Berks
ABSTRACT

Alkaline phosphatases play a crucial role in phosphate acquisition by microorganisms. To expand our understanding of catalysis by this class of enzymes, we have determined the structure of the widely occurring microbial alkaline phosphatase PhoX. The enzyme contains a complex active-site cofactor comprising two antiferromagnetically coupled ferric iron ions (Fe(3+)), three calcium ions (Ca(2+)), and an oxo group bridging three of the metal ions. Notably, the main part of the cofactor resembles synthetic oxide-centered triangular metal complexes. Structures of PhoX-ligand complexes reveal how the active-site metal ions bind substrate and implicate the cofactor oxo group in the catalytic mechanism. The presence of iron in PhoX raises the possibility that iron bioavailability limits microbial phosphate acquisition.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Iron, chips, 99.98% trace metals basis
Iron, IRMM®, certified reference material, 0.5 mm wire
Sigma-Aldrich
Iron, foil, thickness 0.1 mm, ≥99.9% trace metals basis
Sigma-Aldrich
Iron, powder, −325 mesh, 97%
Sigma-Aldrich
Iron, puriss. p.a., carbonyl-Iron powder, low in magnesium and manganese compounds, ≥99.5% (RT)
Iron, tube, 1000mm, outside diameter 12.0mm, inside diameter 9mm, wall thickness 1.5mm, annealed, 99.5%
Iron, tube, 1000mm, outside diameter 25.0mm, inside diameter 21mm, wall thickness 2mm, annealed, 99.5%
Iron, tube, 200mm, outside diameter 6.35mm, inside diameter 5.35mm, wall thickness 0.5mm, as drawn, 99.8%
Iron, tube, 500mm, outside diameter 12.7mm, inside diameter 9.5mm, wall thickness 1.6mm, annealed, 99.5%
Iron, tube, 200mm, outside diameter 2.38mm, inside diameter 1.6mm, wall thickness 0.39mm, as drawn, 99.8+%
Iron, tube, 100mm, outside diameter 3.18mm, inside diameter 1.96mm, wall thickness 0.61mm, as drawn, 99.5%
Iron, tube, 200mm, outside diameter 2.0mm, inside diameter 1.6mm, wall thickness 0.2mm, as drawn, 99.5%
Iron, tube, 200mm, outside diameter 10.0mm, inside diameter 7mm, wall thickness 1.5mm, annealed, 99.5%
Iron, tube, 200mm, outside diameter 20.0mm, inside diameter 16mm, wall thickness 2.0mm, annealed, 99.5%
Iron, tube, 500mm, outside diameter 15.0mm, inside diameter 12mm, wall thickness 1.5mm, annealed, 99.5%
Iron, tube, 500mm, outside diameter 3.18mm, inside diameter 1.96mm, wall thickness 0.61mm, as drawn, 99.5%
Iron, tube, 500mm, outside diameter 5.0mm, inside diameter 4.5mm, wall thickness 0.25mm, as drawn, 99.5%
Iron, tube, 200mm, outside diameter 10.6mm, inside diameter 10.0mm, wall thickness 0.3mm, as drawn, 99.8+%
Iron, tube, 100mm, outside diameter 12.7mm, inside diameter 9.5mm, wall thickness 1.6mm, annealed, 99.5%
Iron, tube, 1000mm, outside diameter 12.0mm, inside diameter 10mm, wall thickness 1.0mm, annealed, 99.5%
Iron, tube, 200mm, outside diameter 25.0mm, inside diameter 21mm, wall thickness 2mm, annealed, 99.5%
Iron, tube, 1000mm, outside diameter 8.0mm, inside diameter 5mm, wall thickness 1.5mm, annealed, 99.5%
Iron, tube, 1000mm, outside diameter 5.0mm, inside diameter 4.5mm, wall thickness 0.25mm, as drawn, 99.5%
Iron, tube, 100mm, outside diameter 10.0mm, inside diameter 7mm, wall thickness 1.5mm, annealed, 99.5%
Iron, tube, 500mm, outside diameter 1.0mm, inside diameter 0.8mm, wall thickness 0.1mm, as drawn, 99.5%
Iron, tube, 500mm, outside diameter 1.65mm, inside diameter 0.89mm, wall thickness 0.38mm, as drawn, 99.8+%
Iron, tube, 200mm, outside diameter 12.0mm, inside diameter 10mm, wall thickness 1.0mm, annealed, 99.5%
Iron, tube, 1000mm, outside diameter 3.2mm, inside diameter 2.2mm, wall thickness 0.5mm, as drawn, 99.8%
Iron, tube, 500mm, outside diameter 6.35mm, inside diameter 5.35mm, wall thickness 0.5mm, as drawn, 99.8%
Iron, tube, 200mm, outside diameter 3.18mm, inside diameter 1.96mm, wall thickness 0.61mm, as drawn, 99.5%