PplD is a de-N-acetylase of the cell wall linkage unit of streptococcal rhamnopolysaccharides

Jeffrey S. Rush; Prakash Parajuli; Alessandro Ruda; Jian Li; Amol Arunrao Pohane; Svetlana Zamakhaeva; Mohammad M. Rahman; Jennifer C. Chang; Artemis Gogos; Cameron W. Kenner; Gérard Lambeau; Michael J. Federle; Konstantin V. Korotkov; Göran Widmalm; Natalia Korotkova

doi:10.1038/s41467-022-28257-0

PplD is a de-N-acetylase of the cell wall linkage unit of streptococcal rhamnopolysaccharides

Jeffrey S. Rush
, Prakash Parajuli
, Alessandro Ruda
, Jian Li
, Amol Arunrao Pohane
, Svetlana Zamakhaeva
, Mohammad M. Rahman
, Jennifer C. Chang
, Artemis Gogos
, Cameron W. Kenner
, Gérard Lambeau
, Michael J. Federle
, Konstantin V. Korotkov
, Göran Widmalm
, Natalia Korotkova

Research output: Contribution to journal › Article › peer-review

19 Scopus citations

Abstract

The cell wall of the human bacterial pathogen Group A Streptococcus (GAS) consists of peptidoglycan decorated with the Lancefield group A carbohydrate (GAC). GAC is a promising target for the development of GAS vaccines. In this study, employing chemical, compositional, and NMR methods, we show that GAC is attached to peptidoglycan via glucosamine 1-phosphate. This structural feature makes the GAC-peptidoglycan linkage highly sensitive to cleavage by nitrous acid and resistant to mild acid conditions. Using this characteristic of the GAS cell wall, we identify PplD as a protein required for deacetylation of linkage N-acetylglucosamine (GlcNAc). X-ray structural analysis indicates that PplD performs catalysis via a modified acid/base mechanism. Genetic surveys in silico together with functional analysis indicate that PplD homologs deacetylate the polysaccharide linkage in many streptococcal species. We further demonstrate that introduction of positive charges to the cell wall by GlcNAc deacetylation protects GAS against host cationic antimicrobial proteins.

Original language	English
Article number	590
Journal	Nature Communications
Volume	13
Issue number	1
DOIs	https://doi.org/10.1038/s41467-022-28257-0
State	Published - Dec 2022

Bibliographical note

Publisher Copyright:
© 2022, The Author(s).

Funding

The authors thank Dr. John F. Timoney (University of Kentucky) for providing S. equi, Dr. Jeffrey M. Bosken, and Dr. Edward D. Hall (University of Kentucky) for the use of the Thermo Fisher Scientific GC-Mass Spectrometer, Dr. Catalina Velez-Ortega (University of Kentucky) for the access to Leica SP8 confocal microscope, and Dr. Peter H. Spielmann (University of Kentucky) for helpful discussions. This work was supported by NIH grants R01 AI143690 from the NIAID and R01 DE028916 from the NIDCR and (to N.K.), the Swedish Research Council (no. 2017-03703) and The Knut and Alice Wal-lenberg Foundation (to G.W.). The Swedish NMR Centre at University of Gothenburg is acknowledged for support. Use of the Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, is supported by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences under Contract No. DE-AC02-76SF00515. The SSRL Structural Molecular Biology Program is supported by the DOE Office of Biological and Environmental Research, and by the National Institutes of Health, National Institute of General Medical Sciences (P30GM133894). The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Funders	Funder number
Knut and Alice Wal-lenberg Foundation
National Institutes of Health (NIH)
U.S. Department of Energy
National Institute of General Medical Sciences DP2GM119177 Sophie Dumont National Institute of General Medical Sciences	P30GM133894
National Institute of Allergy and Infectious F32-AI286447 Cydney N. Johnson Diseases National Institute of Allergy and Infectious R01AI168214 Jason W. Rosch Diseases National Institute of Allergy and Infectious P30 Cydney N. Johnson Diseases National Institute of Allergy and Infectious R00-AI166116 Christopher D. Radka Diseases National Institute of Allergy and Infectious T32-AI106700 Cydney N. Johnson Diseases National Institute of Allergy and Infectious R01AI192221 Jason W. Rosch Diseases National Inst...
National Institute of Dental and Craniofacial Research	R01DE028916
Office of Science Programs
DOE Basic Energy Sciences	DE-AC02-76SF00515
Biological and Environmental Research
Division of Intramural Research, National Institute of Allergy and Infectious Diseases	R01 AI143690
Knut och Alice Wallenbergs Stiftelse
Vetenskapsrådet	2017-03703

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

ASJC Scopus subject areas

General Chemistry
General Biochemistry, Genetics and Molecular Biology
General
General Physics and Astronomy

Access to Document

10.1038/s41467-022-28257-0

Cite this

Rush, J. S., Parajuli, P., Ruda, A., Li, J., Pohane, A. A., Zamakhaeva, S., Rahman, M. M., Chang, J. C., Gogos, A., Kenner, C. W., Lambeau, G., Federle, M. J., Korotkov, K. V., Widmalm, G., & Korotkova, N. (2022). PplD is a de-N-acetylase of the cell wall linkage unit of streptococcal rhamnopolysaccharides. Nature Communications, 13(1), Article 590. https://doi.org/10.1038/s41467-022-28257-0

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abstract = "The cell wall of the human bacterial pathogen Group A Streptococcus (GAS) consists of peptidoglycan decorated with the Lancefield group A carbohydrate (GAC). GAC is a promising target for the development of GAS vaccines. In this study, employing chemical, compositional, and NMR methods, we show that GAC is attached to peptidoglycan via glucosamine 1-phosphate. This structural feature makes the GAC-peptidoglycan linkage highly sensitive to cleavage by nitrous acid and resistant to mild acid conditions. Using this characteristic of the GAS cell wall, we identify PplD as a protein required for deacetylation of linkage N-acetylglucosamine (GlcNAc). X-ray structural analysis indicates that PplD performs catalysis via a modified acid/base mechanism. Genetic surveys in silico together with functional analysis indicate that PplD homologs deacetylate the polysaccharide linkage in many streptococcal species. We further demonstrate that introduction of positive charges to the cell wall by GlcNAc deacetylation protects GAS against host cationic antimicrobial proteins.",

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AU - Gogos, Artemis

AU - Kenner, Cameron W.

AU - Lambeau, Gérard

AU - Federle, Michael J.

AU - Korotkov, Konstantin V.

AU - Widmalm, Göran

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PplD is a de-N-acetylase of the cell wall linkage unit of streptococcal rhamnopolysaccharides

Abstract

Bibliographical note

Funding

UN SDGs

ASJC Scopus subject areas

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