TY - JOUR

T1 - 1-deoxy-D-xylulose-5-phosphate synthase from Pseudomonas aeruginosa and Klebsiella pneumoniae reveals conformational changes upon cofactor binding

AU - Hamid, Rawia

AU - Adam, Sebastian

AU - Lacour, Antoine

AU - Monjas, Leticia

AU - Köhnke, Jesko

AU - Hirsch, Anna K.H.

N1 - Funding Information: The authors would like to thank Dr Nicolas Frank and Dr Timo Risch for native MS and intact MS measurements. We acknowledge the Paul Scherrer Institute, Villigen, Switzerland for the provision of synchrotron radiation beamtime at beamline X06DA-PXIII and X06SA-PXI of the SLS and the European Synchrotron Radiation Facility (ESRF), Grenoble, France for provision of beamline ID23 to 1. A. K. H. H. R. H. and S. A. conceptualization; R. H. and S. A. writing–original draft; A. L. and L. M. synthesis. A. K. H. H. R. H. S. A. A. L. L. M. and J. K. writing–review and editing; A. K. H. H. supervision. The Helmholtz Association's Initiative and Networking Fund (to A. K. H. H.) and the Schlumberger Foundation faculty for the future (FFTF) (to R. H.) funded this work.

PY - 2023/9

Y1 - 2023/9

N2 - The ESKAPE bacteria are the six highly virulent and antibiotic-resistant pathogens that require the most urgent attention for the development of novel antibiotics. Detailed knowledge of target proteins specific to bacteria is essential to develop novel treatment options. The methylerythritol-phosphate (MEP) pathway, which is absent in humans, represents a potentially valuable target for the development of novel antibiotics. Within the MEP pathway, the enzyme 1-deoxy-D-xylulose-5-phosphate synthase (DXPS) catalyzes a crucial, rate-limiting first step and a branch point in the biosynthesis of the vitamins B1 and B6. We report the high-resolution crystal structures of DXPS from the important ESKAPE pathogens Pseudomonas aeruginosa and Klebsiella pneumoniae in both the co-factor-bound and the apo forms. We demonstrate that the absence of the cofactor thiamine diphosphate results in conformational changes that lead to disordered loops close to the active site that might be important for the design of potent DXPS inhibitors. Collectively, our results provide important structural details that aid in the assessment of DXPS as a potential target in the ongoing efforts to combat antibiotic resistance.

AB - The ESKAPE bacteria are the six highly virulent and antibiotic-resistant pathogens that require the most urgent attention for the development of novel antibiotics. Detailed knowledge of target proteins specific to bacteria is essential to develop novel treatment options. The methylerythritol-phosphate (MEP) pathway, which is absent in humans, represents a potentially valuable target for the development of novel antibiotics. Within the MEP pathway, the enzyme 1-deoxy-D-xylulose-5-phosphate synthase (DXPS) catalyzes a crucial, rate-limiting first step and a branch point in the biosynthesis of the vitamins B1 and B6. We report the high-resolution crystal structures of DXPS from the important ESKAPE pathogens Pseudomonas aeruginosa and Klebsiella pneumoniae in both the co-factor-bound and the apo forms. We demonstrate that the absence of the cofactor thiamine diphosphate results in conformational changes that lead to disordered loops close to the active site that might be important for the design of potent DXPS inhibitors. Collectively, our results provide important structural details that aid in the assessment of DXPS as a potential target in the ongoing efforts to combat antibiotic resistance.

KW - 1-deoxy-D-xylulose 5-phosphate synthase

KW - conformational changes

KW - DXPS

KW - Klebsiella pneumonia

KW - Pseudomonas aeruginosa

KW - X-ray crystallography

UR - http://www.scopus.com/inward/record.url?scp=85169906147&partnerID=8YFLogxK

U2 - 10.1016/j.jbc.2023.105152

DO - 10.1016/j.jbc.2023.105152

M3 - Article

C2 - 37567475

AN - SCOPUS:85169906147

VL - 299

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 9

M1 - 105152

ER -