TY - JOUR
T1 - Structural and mechanistic insights into C-P bond hydrolysis by phosphonoacetate hydrolase
AU - Agarwal, Vinayak
AU - Borisova, Svetlana A.
AU - Metcalf, William W.
AU - Van Der Donk, Wilfred A.
AU - Nair, Satish K.
N1 - Funding Information:
This work was supported by the National Institutes of Health (grant GM PO1 GM077596) and the Howard Hughes Medical Institute. Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the National Institute of General Medical Sciences, National Institutes of Health, or Howard Hughes Medical Institute. The authors thank Drs. Keith Brister, Spencer Anderson, and Joseph Brunzelle at LS-CAT (23-ID at Argonne National Laboratories, APS) for facilitating crystallographic data collection. The authors thank Prof. John Gerlt for critical reading of this manuscript.
PY - 2011/10/28
Y1 - 2011/10/28
N2 - Bacteria have evolved pathways to metabolize phosphonates as a nutrient source for phosphorus. In Sinorhizobium meliloti 1021, 2-aminoethylphosphonate is catabolized to phosphonoacetate, which is converted to acetate and inorganic phosphate by phosphonoacetate hydrolase (PhnA). Here we present detailed biochemical and structural characterization of PhnA that provides insights into the mechanism of C-P bond cleavage. The 1.35 resolution crystal structure reveals a catalytic core similar to those of alkaline phosphatases and nucleotide pyrophosphatases but with notable differences, such as a longer metal-metal distance. Detailed structure-guided analysis of active site residues and four additional cocrystal structures with phosphonoacetate substrate, acetate, phosphonoformate inhibitor, and a covalently bound transition state mimic provide insight into active site features that may facilitate cleavage of the C-P bond. These studies expand upon the array of reactions that can be catalyzed by enzymes of the alkaline phosphatase superfamily.
AB - Bacteria have evolved pathways to metabolize phosphonates as a nutrient source for phosphorus. In Sinorhizobium meliloti 1021, 2-aminoethylphosphonate is catabolized to phosphonoacetate, which is converted to acetate and inorganic phosphate by phosphonoacetate hydrolase (PhnA). Here we present detailed biochemical and structural characterization of PhnA that provides insights into the mechanism of C-P bond cleavage. The 1.35 resolution crystal structure reveals a catalytic core similar to those of alkaline phosphatases and nucleotide pyrophosphatases but with notable differences, such as a longer metal-metal distance. Detailed structure-guided analysis of active site residues and four additional cocrystal structures with phosphonoacetate substrate, acetate, phosphonoformate inhibitor, and a covalently bound transition state mimic provide insight into active site features that may facilitate cleavage of the C-P bond. These studies expand upon the array of reactions that can be catalyzed by enzymes of the alkaline phosphatase superfamily.
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U2 - 10.1016/j.chembiol.2011.07.019
DO - 10.1016/j.chembiol.2011.07.019
M3 - Article
C2 - 22035792
AN - SCOPUS:80055084842
SN - 1074-5521
VL - 18
SP - 1230
EP - 1240
JO - Chemistry and Biology
JF - Chemistry and Biology
IS - 10
ER -