4nu5 | pdb_00004nu5

Crystal Structure of PTDH R301A

spinqualitylabelsall models 4nu5, resolution 2.35Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 4nu5 is a 2 chain structure with sequence from "achromobacter_sewerinii"_bergey_et_al._1923 "achromobacter sewerinii" bergey et al. 1923. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Ligands: NAD Related: 4nu6 Gene: ptxD ("Achromobacter sewerinii" Bergey et al. 1923) Activity: Phosphonate dehydrogenase, with EC number 1.20.1.1 Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function [PTXD_PSEST] Catalyzes phosphite (phosphonate) oxidation. Publication Abstract from PubMed Phosphite dehydrogenase (PTDH) catalyzes the NAD(+)-dependent oxidation of phosphite to phosphate. This reaction requires the deprotonation of a water nucleophile for attack on phosphite. A crystal structure was recently solved that identified Arg301 as a potential base given its proximity and orientation to the substrates and a water molecule within the active site. Mutants of this residue showed its importance for efficient catalysis, with about a 100-fold loss in k cat and substantially increased K m,phosphite for the Ala mutant (R301A). The 2.35 A resolution crystal structure of the R301A mutant with NAD(+) bound shows that removal of the guanidine group renders the active site solvent exposed, suggesting the possibility of chemical rescue of activity. We show that the catalytic activity of this mutant is restored to near wild-type levels by the addition of exogenous guanidinium analogues; Bronsted analysis of the rates of chemical rescue suggests that protonation of the rescue reagent is complete in the transition state of the rate-limiting step. Kinetic isotope effects on the reaction in the presence of rescue agents show that hydride transfer remains at least partially rate-limiting, and inhibition experiments show that K i of sulfite with R301A is approximately 400-fold increased compared to the parent enzyme, similar to the increase in K m for phosphite in this mutant. The results of our experiments indicate that Arg301 plays an important role in phosphite binding as well as catalysis, but that it is not likely to act as an active site base. Chemical rescue and inhibition studies to determine the role of arg301 in phosphite dehydrogenase.,Hung JE, Fogle EJ, Garg N, Chekan JR, Nair SK, van der Donk WA PLoS One. 2014 Jan 31;9(1):e87134. doi: 10.1371/journal.pone.0087134. eCollection, 2014. PMID:24498026 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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