5EVO

Structure of Dehydroascrobate Reductase from Pennisetum Americanum in complex with two non-native ligands, Acetate in the G-site and Glycerol in the H-site

Summary for 5EVO

• Entry DOI: 10.2210/pdb5evo/pdb
• Related: 5EVN 5EVP 5EVQ 5EVR 5EVS 5EVT 5EVU 5EVV 5EVW 5EVX
• Descriptor: Dehydroascorbate reductase, ACETATE ION, GLYCEROL, ... (4 entities in total)
• Functional Keywords: apo(native), dhar, oxidoreductase, non-native ligands, acetate (g-site), glycerol (h-site)
• Biological source: Cenchrus americanus (Pearl millet)
• Total number of polymer chains: 1
• Total formula weight: 23877.34

Authors

Kumar, A.O.,Das, B.K.,Arockiasamy, A. (deposition date: 2015-11-20, release date: 2016-05-04, Last modification date: 2024-03-20)

Primary citation

Krishna Das, B.,Kumar, A.,Maindola, P.,Mahanty, S.,Jain, S.K.,Reddy, M.K.,Arockiasamy, A.
Non-native ligands define the active site of Pennisetum glaucum (L.) R. Br dehydroascorbate reductase.
Biochem.Biophys.Res.Commun., 473:1152-1157, 2016

PubMed Abstract: Dehydroascorbate reductase (DHAR), a member of the glutathione-S-transferase (GST) family, reduces dehydroascorbate (DHA) to ascorbate (AsA; Vitamin-C) in a glutathione (GSH)-dependent manner and in doing so, replenishes the critical AsA pool of the cell. To understand the enzyme mechanism in detail, we determined the crystal structure of a plant DHAR from Pennisetum glaucum (PgDHAR) using Iodide-Single Anomalous Dispersion (SAD) and Molecular replacement methods, in two different space groups. Here, we show PgDHAR in complex with two non-native ligands, viz. an acetate bound at the G-site, which resembles the γ-carboxyl moiety of GSH, and a glycerol at the H-site, which shares the backbone of AsA. We also show that, in the absence of bound native substrates, these non-native ligands help define the critical 'hook points' in the DHAR enzyme active site. Further, our data suggest that these non-native ligands can act as the logical bootstrapping points for iterative design of inhibitors/analogs for DHARs.

PubMed: 27067046
DOI: 10.1016/j.bbrc.2016.04.031

Experimental method

X-RAY DIFFRACTION (2.51 Å)