10.1128/AAC.01039-12Antimicrobial Agents and Chemotherapy571168-176AMAC

Basak, S.

Biukovi?, G.

Dick, T.

Gr�ber, G.

Hunke, C.

Manimekalai, M.S.S.

Rao, S.P.S.

Rishikesan, S.

Roessle, M.

PubMed

Subunit ε of bacterial F-ATP synthases plays an important role in regulation by affecting the coupling, catalytic pathway, and inhibition of ATP hydrolysis, achieved through conformational transitions of its C-terminal domain. Here we present the low resolution solution structure of ε of Mycobacterium tuberculosis (Mtε) F-ATP synthase and the NMR structure of its C-terminal segment (Mt103-120), displaying an α-helical region between residues 104-119. The overall length of Mtε (61.6 Å) is significantly shorter compared to other bacterial forms, reflected by the missing C-terminal segment, described to be important in coupling processes via the catalytic β subunit. The surface potential of Mt103-120 reveals highly positive surface charge due to the presence of arginine residues. The unique structural traits of the C-terminus of Mtε make this segment a promising epitope for compounds to bind.

   Using NMR- and fluorescence spectroscopy, we demonstrate that the TB drug TMC207, proposed to bind to the proton translocating c-ring, does also bind to Mtε. W15 has been identified as one of the interacting residues. A model for the interaction of TMC207 with both the ε and the c-ring is presented, showing that TMC207 forms a wedge between the two rotating subunits by interacting with the residues W15 and F50 of ε and the c-ring, respectively. The other two residues, T19 and R37 of ε give the necessary polar interaction to the drug molecule. The new mechanism of TMC207 binding opens a promising platform for the design of antimycobacterials that effictively inhibit the ATP synthase in M. tuberculosis

Biukovic, Goran

Basak, Sandip

Manimekalai, Malathy Sony Subramanian

Rishikeshan, Sankaranarayanana

Roessle, Manfred

Dick, Thomas

Rao, Srinivasa

Hunke, Cornelia

Gruber, Gerhard

The Novartis Repository

Variations of subunit Varepsilon of the Mycobacterium tuberculosis F-1Fo ATP Synthase and Novel Model for Mechanism of Action of the Tuberculosis Drug TMC207.

The subunit ε of bacterial F1FO ATP synthases plays an important regulatory role in coupling and catalysis via conformational transitions of its C-terminal domain. Here we present the first low-resolution solution structure of ε of Mycobacterium tuberculosis (Mtε) F1FO ATP synthase and the nuclear magnetic resonance (NMR) structure of its C-terminal segment (Mtε103–120). Mtε is significantly shorter (61.6 Å) than forms of the subunit in other bacteria, reflecting a shorter C-terminal sequence, proposed to be important in coupling processes via the catalytic β subunit. The C-terminal segment displays an α-helical structure and a highly positive surface charge due to the presence of arginine residues. Using NMR spectroscopy, fluorescence spectroscopy, and mutagenesis, we demonstrate that the new tuberculosis (TB) drug candidate TMC207, proposed to bind to the proton translocating c-ring, also binds to Mtε. A model for the interaction of TMC207 with both ε and the c-ring is presented, suggesting that TMC207 forms a wedge between the two rotating subunits by interacting with the residues W15 and F50 of ε and the c-ring, respectively. T19 and R37 of ε provide the necessary polar interactions with the drug molecule. This new model of the mechanism of TMC207 provides the basis for the design of new drugs targeting the F1FO ATP synthase in M. tuberculosis

Rao, Srinivasa P. S.

Biuković, Goran

Rishikesan, Sankaranarayanan

Grüber, Gerhard

DR-NTU (Digital Repository of NTU)

Antimicrobial Agents and Chemotherapy

Variations of subunit ε of the mycobacterium tuberculosis F1Fo ATP synthase and a novel model for mechanism of action of the tuberculosis drug TMC207

ScholarBank@NUS

Variations of subunit ε of the Mycobacterium tuberculosis F 1FO ATP synthase and a novel model for mechanism of action of the tuberculosis drug TMC207

Digital Repository - Nanyang Technological University

ABSTRACT
          
            The subunit ε of bacterial F
            1
            F
            O
            ATP synthases plays an important regulatory role in coupling and catalysis via conformational transitions of its C-terminal domain. Here we present the first low-resolution solution structure of ε of
            Mycobacterium tuberculosis
            (
            Mt
            ε) F
            1
            F
            O
            ATP synthase and the nuclear magnetic resonance (NMR) structure of its C-terminal segment (
            Mt
            ε
            103–120
            ).
            Mt
            ε is significantly shorter (61.6 Å) than forms of the subunit in other bacteria, reflecting a shorter C-terminal sequence, proposed to be important in coupling processes via the catalytic β subunit. The C-terminal segment displays an α-helical structure and a highly positive surface charge due to the presence of arginine residues. Using NMR spectroscopy, fluorescence spectroscopy, and mutagenesis, we demonstrate that the new tuberculosis (TB) drug candidate TMC207, proposed to bind to the proton translocating
            c
            -ring, also binds to
            Mt
            ε. A model for the interaction of TMC207 with both ε and the
            c
            -ring is presented, suggesting that TMC207 forms a wedge between the two rotating subunits by interacting with the residues W15 and F50 of ε and the
            c
            -ring, respectively. T19 and R37 of ε provide the necessary polar interactions with the drug molecule. This new model of the mechanism of TMC207 provides the basis for the design of new drugs targeting the F
            1
            F
            O
            ATP synthase in
            M. tuberculosis
            .
          </jats:p

Goran Biuković

Sandip Basak

Malathy Sony Subramanian Manimekalai

Sankaranarayanan Rishikesan

Manfred Roessle

Thomas Dick

Srinivasa P. S. Rao

Cornelia Hunke

Gerhard Grüber

Crossref

Variations of Subunit ε of the Mycobacterium tuberculosis F
            <sub>1</sub>
            F
            <sub>o</sub>
            ATP Synthase and a Novel Model for Mechanism of Action of the Tuberculosis Drug TMC207

Variations of subunit ε of the Mycobacterium tuberculosis F 1FO ATP synthase and a novel model for mechanism of action of the tuberculosis drug TMC207

Abstract

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DR-NTU (Digital Repository of NTU)

ScholarBank@NUS

Digital Repository - Nanyang Technological University

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