AFN-1252 is a potent inhibitor of enoyl-ACP reductase from Burkholderia pseudomallei - Crystal structure, mode of action, and biological activity

Krishnamurthy Narasimha Rao, Anirudha Lakshminarasimhan, Sarah Joseph, Swathi U. Lekshmi, Ming Seong Lau, Mohammed Takhi, Kandepu Sreenivas, Sheila Nathan, Rohana Yusof, Noorsaadah Abd. Rahman, Murali Ramachandra, Thomas Antony, Hosahalli Subramanya

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Melioidosis is a tropical bacterial infection caused by Burkholderia pseudomallei (B. pseudomallei; Bpm), a Gram-negative bacterium. Current therapeutic options are largely limited to trimethoprim-sulfamethoxazole and β-lactam drugs, and the treatment duration is about 4 months. Moreover, resistance has been reported to these drugs. Hence, there is a pressing need to develop new antibiotics for Melioidosis. Inhibition of enoyl-ACP reducatase (FabI), a key enzyme in the fatty acid biosynthesis pathway has shown significant promise for antibacterial drug development. FabI has been identified as the major enoyl-ACP reductase present in B. pseudomallei. In this study, we evaluated AFN-1252, a Staphylococcus aureus FabI inhibitor currently in clinical development, for its potential to bind to BpmFabI enzyme and inhibit B. pseudomallei bacterial growth. AFN-1252 stabilized BpmFabI and inhibited the enzyme activity with an IC50 of 9.6 nM. It showed good antibacterial activity against B. pseudomallei R15 strain, isolated from a melioidosis patient (MIC of 2.35 mg/L). X-ray structure of BpmFabI with AFN-1252 was determined at a resolution of 2.3 Å. Complex of BpmFabI with AFN-1252 formed a symmetrical tetrameric structure with one molecule of AFN-1252 bound to each monomeric subunit. The kinetic and thermal melting studies supported the finding that AFN-1252 can bind to BpmFabI independent of cofactor. The structural and mechanistic insights from these studies might help the rational design and development of new FabI inhibitors.

Original languageEnglish
Pages (from-to)832-840
Number of pages9
JournalProtein Science
Volume24
Issue number5
DOIs
Publication statusPublished - 1 May 2015

Fingerprint

Burkholderia pseudomallei
Bioactivity
Oxidoreductases
Crystal structure
Melioidosis
Enzymes
Pharmaceutical Preparations
Lactams
Biosynthesis
Sulfamethoxazole Drug Combination Trimethoprim
Enzyme activity
Gram-Negative Bacteria
Bacterial Infections
Freezing
Inhibitory Concentration 50
Staphylococcus aureus
API 1252
Bacteria
Melting
Fatty Acids

Keywords

  • AFN-1252
  • Burkholderia pseudomallei
  • FabI
  • Melioidosis

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Narasimha Rao, K., Lakshminarasimhan, A., Joseph, S., Lekshmi, S. U., Lau, M. S., Takhi, M., ... Subramanya, H. (2015). AFN-1252 is a potent inhibitor of enoyl-ACP reductase from Burkholderia pseudomallei - Crystal structure, mode of action, and biological activity. Protein Science, 24(5), 832-840. https://doi.org/10.1002/pro.2655

AFN-1252 is a potent inhibitor of enoyl-ACP reductase from Burkholderia pseudomallei - Crystal structure, mode of action, and biological activity. / Narasimha Rao, Krishnamurthy; Lakshminarasimhan, Anirudha; Joseph, Sarah; Lekshmi, Swathi U.; Lau, Ming Seong; Takhi, Mohammed; Sreenivas, Kandepu; Nathan, Sheila; Yusof, Rohana; Abd. Rahman, Noorsaadah; Ramachandra, Murali; Antony, Thomas; Subramanya, Hosahalli.

In: Protein Science, Vol. 24, No. 5, 01.05.2015, p. 832-840.

Research output: Contribution to journalArticle

Narasimha Rao, K, Lakshminarasimhan, A, Joseph, S, Lekshmi, SU, Lau, MS, Takhi, M, Sreenivas, K, Nathan, S, Yusof, R, Abd. Rahman, N, Ramachandra, M, Antony, T & Subramanya, H 2015, 'AFN-1252 is a potent inhibitor of enoyl-ACP reductase from Burkholderia pseudomallei - Crystal structure, mode of action, and biological activity', Protein Science, vol. 24, no. 5, pp. 832-840. https://doi.org/10.1002/pro.2655
Narasimha Rao, Krishnamurthy ; Lakshminarasimhan, Anirudha ; Joseph, Sarah ; Lekshmi, Swathi U. ; Lau, Ming Seong ; Takhi, Mohammed ; Sreenivas, Kandepu ; Nathan, Sheila ; Yusof, Rohana ; Abd. Rahman, Noorsaadah ; Ramachandra, Murali ; Antony, Thomas ; Subramanya, Hosahalli. / AFN-1252 is a potent inhibitor of enoyl-ACP reductase from Burkholderia pseudomallei - Crystal structure, mode of action, and biological activity. In: Protein Science. 2015 ; Vol. 24, No. 5. pp. 832-840.
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abstract = "Melioidosis is a tropical bacterial infection caused by Burkholderia pseudomallei (B. pseudomallei; Bpm), a Gram-negative bacterium. Current therapeutic options are largely limited to trimethoprim-sulfamethoxazole and β-lactam drugs, and the treatment duration is about 4 months. Moreover, resistance has been reported to these drugs. Hence, there is a pressing need to develop new antibiotics for Melioidosis. Inhibition of enoyl-ACP reducatase (FabI), a key enzyme in the fatty acid biosynthesis pathway has shown significant promise for antibacterial drug development. FabI has been identified as the major enoyl-ACP reductase present in B. pseudomallei. In this study, we evaluated AFN-1252, a Staphylococcus aureus FabI inhibitor currently in clinical development, for its potential to bind to BpmFabI enzyme and inhibit B. pseudomallei bacterial growth. AFN-1252 stabilized BpmFabI and inhibited the enzyme activity with an IC50 of 9.6 nM. It showed good antibacterial activity against B. pseudomallei R15 strain, isolated from a melioidosis patient (MIC of 2.35 mg/L). X-ray structure of BpmFabI with AFN-1252 was determined at a resolution of 2.3 {\AA}. Complex of BpmFabI with AFN-1252 formed a symmetrical tetrameric structure with one molecule of AFN-1252 bound to each monomeric subunit. The kinetic and thermal melting studies supported the finding that AFN-1252 can bind to BpmFabI independent of cofactor. The structural and mechanistic insights from these studies might help the rational design and development of new FabI inhibitors.",
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