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Cited by 17Bacterial biofilms and quorum sensingPolysaccharides Composition and ApplicationsEscherichia coli research studies

The DOI matches its Crossref record; 1 of the 1 references it cites were found in a registry or an index.

The TPR domain of PgaA is a multifunctional scaffold that binds PNAG and modulates PgaB-dependent polymer processing

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Pfoh, Roland, Subramanian, Adithya S., Huang, Jingjing, Little, Dustin J., Forman, Adam, DiFrancesco, Benjamin R., Balouchestani-Asli, Negar, Kitova, Elena N., Klassen, John S., Pomès, Régis, Nitz, Mark, Howell, P. Lynne·2022·PLOS Pathogens
DOI 10.1371/journal.ppat.1010750PMC PMC9384988

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Abstract

The synthesis of exopolysaccharides as biofilm matrix components by pathogens is a crucial factor for chronic infections and antibiotic resistance. Many periplasmic proteins involved in polymer processing and secretion in Gram-negative synthase dependent exopolysaccharide biosynthetic systems have been individually characterized. The operons responsible for the production of PNAG, alginate, cellulose and the Pel polysaccharide each contain a gene that encodes an outer membrane associated tetratricopeptide repeat (TPR) domain containing protein. While the TPR domain has been shown to bind other periplasmic proteins, the functional consequences of these interactions for the polymer remain poorly understood. Herein, we show that the C-terminal TPR region of PgaA interacts with the de- N- acetylase domain of PgaB, and increases its deacetylase activity. Additionally, we found that when the two proteins form a complex, the glycoside hydrolase activity of PgaB is also increased. To better understand structure-function relationships we determined the crystal structure of a stable TPR module, which has a conserved groove formed by three repeat motifs. Tryptophan quenching, mass spectrometry analysis and molecular dynamics simulation studies suggest that the crystallized TPR module can bind PNAG/dPNAG via its electronegative groove on the concave surface, and potentially guide the polymer through the periplasm towards the porin for export. Our results suggest a scaffolding role for the TPR domain that combines PNAG/dPNAG translocation with the modulation of its chemical structure by PgaB.

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Metadata

Authors
Pfoh, R., Subramanian, A. S., Huang, J., Little, D. J., Forman, A., DiFrancesco, B. R., Balouchestani-Asli, N., Kitova, E. N., Klassen, J. S., Pomès, R., Nitz, M., & Howell, P. L.
Affiliations
  • Hospital for Sick Children (CA)
  • University of Toronto (CA)
  • University of Alberta (CA)
Corresponding author
Howell, P. Lynne
ORCID
Venue
PLOS Pathogens
Publisher
Public Library of Science (PLoS)
Published
2022-08-05
Volume
18
Issue
8
Pages
e1010750
ISSN
  • 1553-7366 (ISSN-L)
  • 1553-7374
DOI
10.1371/journal.ppat.1010750
PMC
PMC9384988
OpenAlex
W4289973436
Cited by
17
References
1
Citations by year
2026: 2 · 2025: 6 · 2024: 6 · 2023: 3
FWCI
1.22
Type
journal article
Language
en
Update notices
new_version (2022-08-17)
Open access
Licence
Locations
  • PLoS Pathogens (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • OSTI OAI (U.S. Department of Energy Office of Scientific and Technical Information) (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Canadian Institutes of Health Research (MOP 43998 and FDN154327, Graduate scholarship)
  • Natural Sciences and Engineering Research Council of Canada (Discovery Grant, Discovery grant, Graduate Scholarship)
  • Canada Excellence Research Chairs, Government of Canada (Tier I)
  • Hospital for Sick Children (Student Scholarship Program, Graduate Student Program)
  • Department of Medicine, University of Toronto (Graduate scholarship)
  • Province of Ontario (Ontario Graduate Studentship)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bacterial biofilms and quorum sensing
  • Life Sciences › Agricultural and Biological Sciences › Food Science › Polysaccharides Composition and Applications
  • Life Sciences › Immunology and Microbiology › Microbiology › Escherichia coli research studies
Keywords
tetratricopeptide repeat domain, poly-N-acetylglucosamine, exopolysaccharide biosynthesis, protein crystal structure, tryptophan quenching, polymer translocation, biofilm matrix, Pseudomonas aeruginosa, chronic infection
MeSH terms
Tetratricopeptide Repeat, Amidohydrolases, Polymers, Biofilms, Periplasmic Proteins

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