Structural Insights into the Quinolone Resistance Mechanism of Mycobacterium tuberculosis DNA Gyrase

ABSTRACTFluoroquinolone (FQ) resistance is emerging inMycobacterium tuberculosis. The main mechanism of FQ resistance is amino acid substitution within the quinolone resistance-determining region (QRDR) of the GyrA subunit of DNA gyrase, the sole FQ target inM. tuberculosis. However, substitutions in GyrB whose implication in FQ resistance is unknown are increasingly being reported. The present study clarified the role of four GyrB substitutions identified inM. tuberculosisclinical strains, two located in the QRDR (D500A and N538T) and two outside the QRDR (T539P and E540V), in FQ resistance. We measured FQ MICs and also DNA gyrase inhibition by FQs in order to unequivocally clarify the role of these mutations in FQ resistance. Wild-type GyrA, wild-type GyrB, and mutant GyrB subunits produced from engineeredgyrBalleles by mutagenesis were overexpressed inEscherichia coli, purified to homogeneity, and used to reconstitute highly active gyrase complexes. MICs and DNA gyrase inhibition were determined for moxifloxacin, gatifloxacin, ofloxacin, levofloxacin, and enoxacin. All these substitutions are clearly implicated in FQ resistance, underlining the presence of a hot spot region housing most of the GyrB substitutions implicated in FQ resistance (residues NTE, 538 to 540). These findings help us to refine the definition of GyrB QRDR, which is extended to positions 500 to 540.

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Mutagenesis in the α3α4 GyrA Helix and in the Toprim Domain of GyrB Refines the Contribution of Mycobacterium tuberculosis DNA Gyrase to Intrinsic Resistance to Quinolones

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01380-07 ◽

2008 ◽

Vol 52 (8) ◽

pp. 2909-2914 ◽

Cited By ~ 13

Author(s):

Stéphanie Matrat ◽

Alexandra Aubry ◽

Claudine Mayer ◽

Vincent Jarlier ◽

Emmanuelle Cambau

Keyword(s):

Escherichia Coli ◽

Mycobacterium Tuberculosis ◽

Primary Structure ◽

Three Dimensional ◽

Dna Gyrase ◽

Quinolone Resistance ◽

Dimensional Model ◽

Three Dimensional Model ◽

Intrinsic Resistance ◽

E Coli

ABSTRACT The replacement of M74 in GyrA, A83 in GyrA, and R447 in GyrB of Mycobacterium tuberculosis gyrase by their Escherichia coli homologs resulted in active enzymes as quinolone susceptible as the E. coli gyrase. This demonstrates that the primary structure of gyrase determines intrinsic quinolone resistance and was supported by a three-dimensional model of N-terminal GyrA.

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Structure-Based Virtual Screening of Benzaldehyde Thiosemicarbazone Derivatives against DNA Gyrase B of Mycobacterium tuberculosis

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2021/6140378 ◽

2021 ◽

Vol 2021 ◽

pp. 1-11

Author(s):

Andiyappan Kistan ◽

Balakrishnan Anna Benedict ◽

Sundaramoorthy Vasanthan ◽

Alphonse PremKumar ◽

Malathi Kullappan ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Resistance Mechanism ◽

Dna Gyrase ◽

The Novel ◽

Extrinsic Factors ◽

Replication Process ◽

Antibiotic Resistant ◽

Compound 21 ◽

Benzaldehyde Thiosemicarbazone ◽

Intrinsic And Extrinsic Factors

Emergence of antibiotic-resistant Mycobacterium tuberculosis (M. tuberculosis) restricts the availability of drugs for the treatment of tuberculosis, which leads to the increased morbidity and mortality of the disease worldwide. There are many intrinsic and extrinsic factors that have been reported for the resistance mechanism. To overcome such mechanisms, chemically synthesized benzaldehyde thiosemicarbazone derivatives were screened against M. tuberculosis to find potential inhibitor for tuberculosis. Such filtering process resulted in compound 13, compound 21, and compound 20 as the best binding energy compounds against DNA gyrase B, an important protein in the replication process. The ADMET prediction has shown the oral bioavailability of the novel compounds.

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Structural investigation of inhibitor designs targeting 3-dehydroquinate dehydratase from the shikimate pathway of Mycobacterium tuberculosis

Biochemical Journal ◽

10.1042/bj20110002 ◽

2011 ◽

Vol 436 (3) ◽

pp. 729-739 ◽

Cited By ~ 23

Author(s):

Marcio V. B. Dias ◽

William C. Snee ◽

Karen M. Bromfield ◽

Richard J. Payne ◽

Satheesh K. Palaninathan ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Drug Targets ◽

Structural Investigation ◽

Catalytic Mechanism ◽

Shikimate Pathway ◽

Structural Rearrangement ◽

Competitive Inhibitors ◽

Structural Insights ◽

Potential Drug Targets

The shikimate pathway is essential in Mycobacterium tuberculosis and its absence from humans makes the enzymes of this pathway potential drug targets. In the present paper, we provide structural insights into ligand and inhibitor binding to 3-dehydroquinate dehydratase (dehydroquinase) from M. tuberculosis (MtDHQase), the third enzyme of the shikimate pathway. The enzyme has been crystallized in complex with its reaction product, 3-dehydroshikimate, and with six different competitive inhibitors. The inhibitor 2,3-anhydroquinate mimics the flattened enol/enolate reaction intermediate and serves as an anchor molecule for four of the inhibitors investigated. MtDHQase also forms a complex with citrazinic acid, a planar analogue of the reaction product. The structure of MtDHQase in complex with a 2,3-anhydroquinate moiety attached to a biaryl group shows that this group extends to an active-site subpocket inducing significant structural rearrangement. The flexible extensions of inhibitors designed to form π-stacking interactions with the catalytic Tyr24 have been investigated. The high-resolution crystal structures of the MtDHQase complexes provide structural evidence for the role of the loop residues 19–24 in MtDHQase ligand binding and catalytic mechanism and provide a rationale for the design and efficacy of inhibitors.

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Structural Insights of LspA, from Mycobacterium Tuberculosis, using Solid State NMR Spectroscopy

Biophysical Journal ◽

10.1016/j.bpj.2013.11.348 ◽

2014 ◽

Vol 106 (2) ◽

pp. 48a-49a

Author(s):

E. Vindana Ekanayake ◽

Huajun Qin ◽

Ivan Hung ◽

Timothy A. Cross

Keyword(s):

Mycobacterium Tuberculosis ◽

Solid State ◽

Nmr Spectroscopy ◽

Solid State Nmr ◽

Solid State Nmr Spectroscopy ◽

Structural Insights

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Quinolones: Mechanism, Lethality and Their Contributions to Antibiotic Resistance

Molecules ◽

10.3390/molecules25235662 ◽

2020 ◽

Vol 25 (23) ◽

pp. 5662

Author(s):

Natassja G. Bush ◽

Isabel Diez-Santos ◽

Lauren R. Abbott ◽

Anthony Maxwell

Keyword(s):

Cell Death ◽

Antibiotic Resistance ◽

Dna Gyrase ◽

Mechanisms Of Action ◽

Quinolone Resistance ◽

Topoisomerase Iv ◽

Bacterial Enzymes ◽

Dna Topoisomerase ◽

Quinolone Antibiotics ◽

Dna Complex

Fluoroquinolones (FQs) are arguably among the most successful antibiotics of recent times. They have enjoyed over 30 years of clinical usage and become essential tools in the armoury of clinical treatments. FQs target the bacterial enzymes DNA gyrase and DNA topoisomerase IV, where they stabilise a covalent enzyme-DNA complex in which the DNA is cleaved in both strands. This leads to cell death and turns out to be a very effective way of killing bacteria. However, resistance to FQs is increasingly problematic, and alternative compounds are urgently needed. Here, we review the mechanisms of action of FQs and discuss the potential pathways leading to cell death. We also discuss quinolone resistance and how quinolone treatment can lead to resistance to non-quinolone antibiotics.

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Mutation in the DNA Gyrase A Gene ofEscherichia coli That Expands the Quinolone Resistance-Determining Region

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.45.8.2378-2380.2001 ◽

2001 ◽

Vol 45 (8) ◽

pp. 2378-2380 ◽

Cited By ~ 66

Author(s):

S. Marvin Friedman ◽

Tao Lu ◽

Karl Drlica

Keyword(s):

Escherichia Coli ◽

Amino Acids ◽

Amino Acid ◽

Dna Gyrase ◽

Ofescherichia Coli ◽

Quinolone Resistance ◽

Gyrase A

ABSTRACT In three Escherichia coli mutants, a change (Ala-51 to Val) in the gyrase A protein outside the standard quinolone resistance-determining region (QRDR) lowered the level of quinolone susceptibility more than changes at amino acids 67, 82, 84, and 106 did. Revision of the QRDR to include amino acid 51 is indicated.

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