The lytic transglycosylase, LtgG, controls cell morphology and virulence in Burkholderia pseudomallei

DOI: 10.1038/s41598-019-47483-z

Authors: Christopher H. Jenkins (University of Leicester; Defence Science and Technology Laboratory) , Russell Wallis (University of Leicester) , Natalie Allcock (University of Leicester) , Kay B. Barnes (Defence Science and Technology Laboratory) , Mark I. Richards (Defence Science and Technology Laboratory) , Joss M. Auty (University of Leicester) , Edouard E. Galyov (University of Leicester) , Sarah V. Harding (Defence Science and Technology Laboratory; University of Leicester) , Galina V. Mukamolova (University of Leicester)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Scientific Reports , VOL 9

State: Published (Approved)
Published: July 2019
Diamond Proposal Number(s): 10369

Abstract: Burkholderia pseudomallei is the causative agent of the tropical disease melioidosis. Its genome encodes an arsenal of virulence factors that allow it, when required, to switch from a soil dwelling bacterium to a deadly intracellular pathogen. With a high intrinsic resistance to antibiotics and the ability to overcome challenges from the host immune system, there is an increasing requirement for new antibiotics and a greater understanding into the molecular mechanisms of B. pseudomallei virulence and dormancy. The peptidoglycan remodeling enzymes, lytic transglycosylases (Ltgs) are potential targets for such new antibiotics. Ltgs cleave the glycosidic bonds within bacterial peptidoglycan allowing for the insertion of peptidoglycan precursors during cell growth and division, and cell membrane spanning structures such as flagella and secretion systems. Using bioinformatic analysis we have identified 8 putative Ltgs in B. pseudomallei K96243. We aimed to investigate one of these Ltgs, LtgG (BPSL3046) through the generation of deletion mutants and biochemical analysis. We have shown that LtgG is a key contributor to cellular morphology, division, motility and virulence in BALB/c mice. We have determined the crystal structure of LtgG and have identified various amino acids likely to be important in peptidoglycan binding and catalytic activity. Recombinant protein assays and complementation studies using LtgG containing a site directed mutation in aspartate 343, confirmed the essentiality of this amino acid in the function of LtgG.

Journal Keywords: Bacterial physiology; Enzymes

Diamond Keywords: Bacteria; Enzymes; Melioidosis

Subject Areas: Biology and Bio-materials, Medicine


Instruments: I03-Macromolecular Crystallography

Added On: 14/08/2019 08:56

Documents:
s41598-019-47483-z.pdf

Discipline Tags:

Pathogens Infectious Diseases Disease in the Developing World Health & Wellbeing Structural biology Drug Discovery Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)