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

9 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)
Michigan State University-U.S. Department of Energy (MSU-DOE) Plant Research Laboratory
National Institute of General Medical Sciences	P30GM133894
National Institute of Allergy and Infectious Diseases
National Institute of Dental and Craniofacial Research	R01DE028916
Office of Science Programs
Office of 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

ASJC Scopus subject areas

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

UN SDGs

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

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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 - Pohane, Amol Arunrao

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AU - Kenner, Cameron W.

AU - Lambeau, Gérard

AU - Federle, Michael J.

AU - Korotkov, Konstantin V.

AU - Widmalm, Göran

AU - Korotkova, Natalia

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

Abstract

Bibliographical note

Funding

ASJC Scopus subject areas

UN SDGs

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