5J62

FMN-dependent Nitroreductase (CDR20291_0684) from Clostridium difficile R20291

Summary for 5J62

• Entry DOI: 10.2210/pdb5j62/pdb
• Related: 5J6C
• Descriptor: Putative reductase, FLAVIN MONONUCLEOTIDE, GLYCEROL, ... (5 entities in total)
• Functional Keywords: nitroreductase, hypervirulent, clostridium difficile, r20291, metronidazole resistance, oxidoreductase
• Biological source: Peptoclostridium difficile (strain R20291)
• Total number of polymer chains: 2
• Total formula weight: 53242.42

Authors

Wang, B.,Powell, S.M.,Hessami, N.,Najar, F.Z.,Thomas, L.M.,West, A.H.,Karr, E.A.,Richter-Addo, G.B. (deposition date: 2016-04-04, release date: 2016-09-21, Last modification date: 2023-09-27)

Primary citation

Wang, B.,Powell, S.M.,Hessami, N.,Najar, F.Z.,Thomas, L.M.,Karr, E.A.,West, A.H.,Richter-Addo, G.B.
Crystal structures of two nitroreductases from hypervirulent Clostridium difficile and functionally related interactions with the antibiotic metronidazole.
Nitric Oxide, 60:32-39, 2016

PubMed Abstract: Nitroreductases (NRs) are flavin mononucleotide (FMN)-dependent enzymes that catalyze the biotransformation of organic nitro compounds (RNO; R = alkyl, aryl) to the nitroso RN=O, hydroxylamino RNHOH, or amine RNH derivatives. Metronidazole (Mtz) is a nitro-containing antibiotic that is commonly prescribed for lower-gut infections caused by the anaerobic bacterium Clostridium difficile. C. difficile infections rank number one among hospital acquired infections, and can result in diarrhea, severe colitis, or even death. Although NRs have been implicated in Mtz resistance of C. difficile, no NRs have been characterized from the hypervirulent R20291 strain of C. difficile. We report the first expression, purification, and three-dimensional X-ray crystal structures of two NRs from the C. difficile R20291 strain. The X-ray crystal structures of the two NRs were solved to 2.1 Å resolution. Their homodimeric structures exhibit the classic NR α+β fold, with each protomer binding one FMN cofactor near the dimer interface. Functional assays demonstrate that these two NRs metabolize Mtz with associated re-oxidation of the proteins. Importantly, these results represent the first isolation and characterization of NRs from the hypervirulent R20291 strain of relevance to organic RNO (e.g., Mtz) metabolism.

PubMed: 27623089
DOI: 10.1016/j.niox.2016.09.003

Experimental method

X-RAY DIFFRACTION (2.15 Å)