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The antibiotic novobiocin binds and activates the ATPase that powers lipopolysaccharide transport

May, Janine M. and Owens, Tristan W. and Mandler, Michael D. and Simpson, Brent W. and Lazarus, Michael B. and Sherman, David J. and Davis, Rebecca M. and Okuda, Suguru and Massefski, Walter and Ruiz, Natividad and Kahne, Daniel (2017) The antibiotic novobiocin binds and activates the ATPase that powers lipopolysaccharide transport. Journal of the American Chemical Society, 139 (48). pp. 17221-17224. ISSN 0002-7863.

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Novobiocin is an orally active antibiotic that inhibits DNA gyrase by binding the ATP-binding site in the ATPase subunit. Although effective against Gram-positive pathogens, novobiocin has limited activity against Gram-negative organisms due to the presence of the lipopolysaccharide-containing outer membrane, which acts as a permeability barrier. Using a novobiocin-sensitive Escherichia coli strain with a leaky outer membrane, we identified a mutant with increased resistance to novobiocin. Unexpectedly, the mutation that increases novobiocin resistance was not found to alter gyrase, but the ATPase that powers lipopolysaccharide (LPS) transport. Co-crystal structures, biochemical, and genetic evidence show novobiocin directly binds this ATPase. Novobiocin does not bind the ATP binding site but rather the interface between the ATPase subunits and the transmembrane subunits of the LPS transporter. This interaction increases the activity of the LPS transporter, which in turn alters the permeability of the outer membrane. We propose that novobiocin will be a useful tool for understanding how ATP hydrolysis is coupled to LPS transport.

Item Type:Article
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URLURL TypeDescription Information
Kahne, Daniel0000-0002-8296-1424
Additional Information:© 2017 American Chemical Society. Received: July 24, 2017; Published: November 14, 2017. This research was supported by the NIH (R01 GM100951 to N.R.; R01 GM066174, U19 AI109764, and R01 AI081059 to D.K.), NSF GRFP (DGE-1144152 to M.D.M.), and the Blavatnik Biomedical Accelerator at Harvard University. We wish to thank the Dana Farber Cancer Institute/Harvard Medical School joint NMR Core. This work used NE-CAT beamlines (GM103403), a Pilatus detector (RR029205), and an Eiger detector (OD021527) at the APS (DE-AC02-06CH11357). The authors declare no competing financial interest.
Funding AgencyGrant Number
NIHR01 GM100951
NIHR01 GM066174
NIHU19 AI109764
NIHR01 AI081059
NSF Graduate Research FellowshipDGE-1144152
Harvard UniversityUNSPECIFIED
Dana Farber Cancer InstituteUNSPECIFIED
Department of Energy (DOE)DE-AC02-06CH11357
Issue or Number:48
Record Number:CaltechAUTHORS:20171115-102801445
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Official Citation:The Antibiotic Novobiocin Binds and Activates the ATPase That Powers Lipopolysaccharide Transport. Janine M. May, Tristan W. Owens, Michael D. Mandler, Brent W. Simpson, Michael B. Lazarus, David J. Sherman, Rebecca M. Davis, Suguru Okuda, Walter Massefski, Natividad Ruiz, and Daniel Kahne. Journal of the American Chemical Society 2017 139 (48), 17221-17224. DOI: 10.1021/jacs.7b07736
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:83221
Deposited By: Tony Diaz
Deposited On:15 Nov 2017 18:38
Last Modified:03 Oct 2019 19:03

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