TY - JOUR
T1 - Phosphate-binding protein from Polaromonas JS666
T2 - Purification, characterization, crystallization and sulfur SAD phasing
AU - Pegos, Vanessa R.
AU - Hey, Louis
AU - Lamirande, Jacob
AU - Pfeffer, Rachel
AU - Lipsh, Rosalie
AU - Amitay, Moshe
AU - Gonzalez, Daniel
AU - Elias, Mikael
N1 - Publisher Copyright:
© International Union of Crystallography, 2017.
PY - 2017/6
Y1 - 2017/6
N2 - Phosphate-binding proteins (PBPs) are key proteins that belong to the bacterial ABC-type phosphate transporters. PBPs are periplasmic (or membrane-anchored) proteins that capture phosphate anions from the environment and release them to the transmembrane transporter. Recent work has suggested that PBPs have evolved for high affinity as well as high selectivity. In particular, a short, unique hydrogen bond between the phosphate anion and an aspartate residue has been shown to be critical for selectivity, yet is not strictly conserved in PBPs. Here, the PBP from Polaromonas JS666 is focused on. Interestingly, this PBP is predicted to harbor different phosphate-binding residues to currently known PBPs. Here, it is shown that the PBP from Polaromonas JS666 is capable of binding phosphate, with a maximal binding activity at pH 8. Its structure is expected to reveal its binding-cleft configuration as well as its phosphate-binding mode. Here, the expression, purification, characterization, crystallization and X-ray diffraction data collection to 1.35 Å resolution of the PBP from Polaromonas JS666 are reported.
AB - Phosphate-binding proteins (PBPs) are key proteins that belong to the bacterial ABC-type phosphate transporters. PBPs are periplasmic (or membrane-anchored) proteins that capture phosphate anions from the environment and release them to the transmembrane transporter. Recent work has suggested that PBPs have evolved for high affinity as well as high selectivity. In particular, a short, unique hydrogen bond between the phosphate anion and an aspartate residue has been shown to be critical for selectivity, yet is not strictly conserved in PBPs. Here, the PBP from Polaromonas JS666 is focused on. Interestingly, this PBP is predicted to harbor different phosphate-binding residues to currently known PBPs. Here, it is shown that the PBP from Polaromonas JS666 is capable of binding phosphate, with a maximal binding activity at pH 8. Its structure is expected to reveal its binding-cleft configuration as well as its phosphate-binding mode. Here, the expression, purification, characterization, crystallization and X-ray diffraction data collection to 1.35 Å resolution of the PBP from Polaromonas JS666 are reported.
KW - Polaromonas JS666
KW - molecular specificity
KW - phosphate ABC transporter
KW - phosphate-binding protein
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U2 - 10.1107/S2053230X17007373
DO - 10.1107/S2053230X17007373
M3 - Article
C2 - 28580922
AN - SCOPUS:85020242909
SN - 1744-3091
VL - 73
SP - 342
EP - 346
JO - Acta Crystallographica Section:F Structural Biology Communications
JF - Acta Crystallographica Section:F Structural Biology Communications
IS - 6
ER -