5TB7

Structure of nitrite reductase AniA from Neisseria gonorrhoeae, space group P212121

Summary for 5TB7

• Entry DOI: 10.2210/pdb5tb7/pdb
• Descriptor: Nitrite reductase, COPPER (II) ION, PHOSPHATE ION, ... (4 entities in total)
• Functional Keywords: ngo1276, oxidoreductase
• Biological source: Neisseria gonorrhoeae (strain ATCC 700825 / FA 1090)
• Total number of polymer chains: 3
• Total formula weight: 109860.16

Authors

Hamza, A.,Williamson, Z.A.,Reed, R.W.,Sikora, A.E.,Korotkov, K.V. (deposition date: 2016-09-11, release date: 2017-02-01, Last modification date: 2023-10-04)

Primary citation

Sikora, A.E.,Mills, R.H.,Weber, J.V.,Hamza, A.,Passow, B.W.,Romaine, A.,Williamson, Z.A.,Reed, R.W.,Zielke, R.A.,Korotkov, K.V.
Peptide Inhibitors Targeting the Neisseria gonorrhoeae Pivotal Anaerobic Respiration Factor AniA.
Antimicrob. Agents Chemother., 61:-, 2017

PubMed Abstract: causes the sexually transmitted infection gonorrhea, which is highly prevalent worldwide and has a major impact on reproductive and neonatal health. The superbug status of necessitates the development of drugs with different mechanisms of action. Here, we focused on targeting the nitrite reductase AniA, which is a pivotal component of anaerobic respiration and biofilm formation. Our studies showed that gonococci expressing AniA containing the altered catalytic residues D137A and H280A failed to grow under anaerobic conditions, demonstrating that the nitrite reductase function is essential. To facilitate the pharmacological targeting of AniA, new crystal structures of AniA were refined to 1.90-Å and 2.35-Å resolutions, and a phage display approach with libraries expressing randomized linear dodecameric peptides or heptameric peptides flanked by a pair of cysteine residues was utilized. Biopanning experiments led to the identification of 29 unique peptides, with 1 of them, C7-3, being identified multiple times. Evaluation of their ability to interact with AniA using enzyme-linked immunosorbent assay and computational docking studies revealed that C7-3 was the most promising inhibitor, binding near the type 2 copper site of the enzyme, which is responsible for interaction with nitrite. Subsequent enzymatic assays and biolayer interferometry with a synthetic C7-3 and its derivatives, C7-3m1 and C7-3m2, demonstrated potent inhibition of AniA. Finally, the MIC value of C7-3 and C7-3m2 against anaerobically grown was 0.6 mM. We present the first peptide inhibitors of AniA, an enzyme that should be further exploited for antigonococcal drug development.

PubMed: 28584144
DOI: 10.1128/AAC.00186-17

Experimental method

X-RAY DIFFRACTION (1.9 Å)