TY - JOUR
T1 - Structure and function of phosphonoacetaldehyde dehydrogenase
T2 - The missing link in phosphonoacetate formation
AU - Agarwal, Vinayak
AU - Peck, Spencer C.
AU - Chen, Jui Hui
AU - Borisova, Svetlana A.
AU - Chekan, Jonathan R.
AU - Van Der Donk, Wilfred A.
AU - Nair, Satish K.
N1 - Funding Information:
We thank Drs. Keith Brister and Joseph Brunzelle at the LS-CAT (Sector-21) beamline at Argonne National Laboratory for assistance with X-ray diffraction data collection and Prof. John A. Gerlt (University of Illinois) for stimulating discussions and S. mutans genomic DNA. Use of the Advanced Photon Source, an Office of Science User Facility operated for the US Department of Energy (DOE) Office of Science by Argonne National Laboratory, was supported by the US DOE under contract no. DE-AC02-06CH11357. Use of the LS-CAT Sector 21 was supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor (grant 085P1000817). NMR spectra were recorded on a 600 MHz instrument purchased with support from NIH S10 RR028833. This work was supported by the National Institutes of Health (GM P01 077596 to W.A.V. and S.K.N.)
PY - 2014/1/16
Y1 - 2014/1/16
N2 - Summary Phosphonates (C-PO32-) have applications as antibiotics, herbicides, and detergents. In some environments, these molecules represent the predominant source of phosphorus, and several microbes have evolved dedicated enzymatic machineries for phosphonate degradation. For example, most common naturally occurring phosphonates can be catabolized to either phosphonoacetaldehyde or phosphonoacetate, which can then be hydrolyzed to generate inorganic phosphate and acetaldehyde or acetate, respectively. The phosphonoacetaldehyde oxidase gene (phnY) links these two hydrolytic processes and provides a previously unknown catabolic mechanism for phosphonoacetate production in the microbial metabolome. Here, we present biochemical characterization of PhnY and high-resolution crystal structures of the apo state, as well as complexes with substrate, cofactor, and product. Kinetic analysis of active site mutants demonstrates how a highly conserved aldehyde dehydrogenase active site has been modified in nature to generate activity with a phosphonate substrate.
AB - Summary Phosphonates (C-PO32-) have applications as antibiotics, herbicides, and detergents. In some environments, these molecules represent the predominant source of phosphorus, and several microbes have evolved dedicated enzymatic machineries for phosphonate degradation. For example, most common naturally occurring phosphonates can be catabolized to either phosphonoacetaldehyde or phosphonoacetate, which can then be hydrolyzed to generate inorganic phosphate and acetaldehyde or acetate, respectively. The phosphonoacetaldehyde oxidase gene (phnY) links these two hydrolytic processes and provides a previously unknown catabolic mechanism for phosphonoacetate production in the microbial metabolome. Here, we present biochemical characterization of PhnY and high-resolution crystal structures of the apo state, as well as complexes with substrate, cofactor, and product. Kinetic analysis of active site mutants demonstrates how a highly conserved aldehyde dehydrogenase active site has been modified in nature to generate activity with a phosphonate substrate.
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U2 - 10.1016/j.chembiol.2013.11.006
DO - 10.1016/j.chembiol.2013.11.006
M3 - Article
C2 - 24361046
AN - SCOPUS:84892668683
SN - 1074-5521
VL - 21
SP - 125
EP - 135
JO - Chemistry and Biology
JF - Chemistry and Biology
IS - 1
ER -