Unraveling the B. pseudomallei Heptokinase WcbL: From Structure to Drug Discovery. Issue 12 (17th December 2015)

• Record Type: Journal Article
• Authors: Vivoli, Mirella; Isupov, Michail N.; Nicholas, Rebecca; Hill, Andrew; Scott, Andrew E.; Kosma, Paul; Prior, Joann L.; Harmer, Nicholas J.
• Is Part Of: Chemistry & biology. Volume 22:Issue 12(2015)
• Page Start: 1622
• Page End: 1632
• Publication Date: 2015-12-17
• Subjects: Biochemistry -- Periodicals; 540
• DOI: 10.1016/j.chembiol.2015.10.015
• Languages: English
• ISSNs: 1074-5521

Abstract

Summary: Gram-negative bacteria utilize heptoses as part of their repertoire of extracellular polysaccharide virulence determinants. Disruption of heptose biosynthesis offers an attractive target for novel antimicrobials. A critical step in the synthesis of heptoses is their 1- O phosphorylation, mediated by kinases such as HldE or WcbL. Here, we present the structure of WcbL from Burkholderia pseudomallei . We report that WcbL operates through a sequential ordered Bi-Bi mechanism, loading the heptose first and then ATP. We show that dimeric WcbL binds ATP anti-cooperatively in the absence of heptose, and cooperatively in its presence. Modeling of WcbL suggests that heptose binding causes an elegant switch in the hydrogen-bonding network, facilitating the binding of a second ATP molecule. Finally, we screened a library of drug-like fragments, identifying hits that potently inhibit WcbL. Our results provide a novel mechanism for control of substrate binding and emphasize WcbL as an attractive anti-microbial target for Gram-negative bacteria. Graphical Abstract: Highlights: The structure of WcbL in complex with ligand analogs is reported WcbL switches its cooperativity toward ATP in the presence of sugars The kinetics and mechanism of WcbL are elucidated Screening of a fragment library identified a potent inhibitor of WcbL Abstract : Vivoli et al. present the structure and mechanism of the bacterial sugar kinase WcbL. WcbL demonstrates unusual kinetics: its cooperativity for ATP switches on binding of the sugar substrate. Screening of a fragment library identified a potent inhibitor of WcbL.