Characterisation of trehalose-6-phosphate phosphatases from bacterial pathogens

Loading...
Thumbnail Image
File version
Accepted Manuscript (AM)
Author(s)
Kim, Jun-Hong
Kim, Ji-Won
Jo, Jiwon
Straub, Jan Hendrik
Cross, Megan
Hofmann, Andreas
Kim, Jeong-Sun
Griffith University Author(s)
Primary Supervisor
Other Supervisors
Editor(s)
Date
2020
Size
File type(s)
Location
License
http://creativecommons.org/licenses/by-nc-nd/4.0/
Abstract

The trehalose biosynthesis pathway has recently received attention for therapeutic intervention combating infectious diseases caused by bacteria, helminths or fungi. Trehalose-6-phosphate phosphatase (TPP) is a key enzyme of the most common trehalose biosynthesis pathway and a particularly attractive target owing to the toxicity of accumulated trehalose-6-phosphate in pathogens. Here, we characterised TPP-like proteins from bacterial pathogens implicated in nosocomial infections in terms of their steady-state kinetics as well as pH- and metal-dependency of their enzymatic activity. Analysis of the steady-state kinetics of recombinantly expressed enzymes from Acinetobacter baumannii, Corynebacterium diphtheriae and Pseudomonas stutzeri yielded similar kinetic parameters as those of other reported bacterial TPPs. In contrast to nematode TPPs, the divalent metal ion appears to be bound only weakly in the active site of bacterial TPPs, allowing the exchange of the resident magnesium ion with other metal ions. Enzymatic activity comparable to the wild-type enzyme was observed for the TPP from P. stutzeri with manganese, cobalt and nickel. Analysis of the enzymatic activity of S. maltophilia TPP active site mutants provides evidence for the involvement of four canonical aspartate residues as well as a strictly conserved histidine residue of TPP-like proteins from bacteria in the enzyme mechanism. That histidine residue is a member of an interconnected network of five conserved residues in the active site of bacterial TPPs which likely constitute one or more functional units, directly or indirectly cooperating to enhance different aspects of the catalytic activity.

Journal Title
Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics
Conference Title
Book Title
Edition
Volume
1869
Issue
2
Thesis Type
Degree Program
School
Publisher link
Patent number
Funder(s)
Grant identifier(s)
Rights Statement
Rights Statement
© 2020 Elsevier. Licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International Licence (http://creativecommons.org/licenses/by-nc-nd/4.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, providing that the work is properly cited.
Item Access Status
Note
Access the data
Related item(s)
Subject
Biological sciences
Catalyis
Cooperativity
Enzyme activity
Halo-acid dehydrogenase
Multi-drug resistance
Persistent link to this record
Citation
Kim, J-H; Kim, J-W; Jo, J; Straub, JH; Cross, M; Hofmann, A; Kim, J-S, Characterisation of trehalose-6-phosphate phosphatases from bacterial pathogens, Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics, 2020, 1869 (2), pp. 140564
Collections