Discovery of glycerol phosphate modification on streptococcal rhamnose polysaccharides

Rebecca J Edgar; Vincent P van Hensbergen; Alessandro Ruda; Andrew G Turner; Pan Deng; Yoann Le Breton; Najib M El-Sayed; Ashton T Belew; Kevin S McIver; Alastair G McEwan; Andrew J Morris; Gérard Lambeau; Mark J Walker; Jeffrey S Rush; Konstantin V Korotkov; Göran Widmalm; Nina M van Sorge; Natalia Korotkova

doi:10.1038/s41589-019-0251-4

Discovery of glycerol phosphate modification on streptococcal rhamnose polysaccharides

Nat Chem Biol. 2019 May;15(5):463-471. doi: 10.1038/s41589-019-0251-4. Epub 2019 Apr 1.

Authors

Rebecca J Edgar^{1

2}, Vincent P van Hensbergen³, Alessandro Ruda⁴, Andrew G Turner⁵, Pan Deng⁶, Yoann Le Breton^{7

8}, Najib M El-Sayed^{7

9}, Ashton T Belew^{7

9}, Kevin S McIver⁷, Alastair G McEwan⁵, Andrew J Morris⁶, Gérard Lambeau¹⁰, Mark J Walker⁵, Jeffrey S Rush¹, Konstantin V Korotkov¹, Göran Widmalm⁴, Nina M van Sorge¹¹, Natalia Korotkova¹²

Affiliations

¹ Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, KY, USA.
² Callaghan Innovation, Gracefield, Lower Hutt, New Zealand.
³ Department of Medical Microbiology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands.
⁴ Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University, Stockholm, Sweden.
⁵ Australian Infectious Diseases Research Centre and School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, Australia.
⁶ Division of Cardiovascular Medicine and the Gill Heart Institute, University of Kentucky, Lexington, KY, USA.
⁷ Department of Cell Biology & Molecular Genetics and Maryland Pathogen Research Institute, University of Maryland, College Park, MD, USA.
⁸ Wound Infections Department, Bacterial Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, MD, USA.
⁹ Center for Bioinformatics and Computational Biology, University of Maryland, College Park, MD, USA.
¹⁰ Université Côte d'Azur, Centre National de la Recherche Scientifique, Institut de Pharmacologie Moléculaire et Cellulaire, Valbonne Sophia Antipolis, France.
¹¹ Department of Medical Microbiology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands. nsorge3@umcutrecht.nl.
¹² Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, KY, USA. nkorotkova@uky.edu.

Abstract

Cell wall glycopolymers on the surface of Gram-positive bacteria are fundamental to bacterial physiology and infection biology. Here we identify gacH, a gene in the Streptococcus pyogenes group A carbohydrate (GAC) biosynthetic cluster, in two independent transposon library screens for its ability to confer resistance to zinc and susceptibility to the bactericidal enzyme human group IIA-secreted phospholipase A₂. Subsequent structural and phylogenetic analysis of the GacH extracellular domain revealed that GacH represents an alternative class of glycerol phosphate transferase. We detected the presence of glycerol phosphate in the GAC, as well as the serotype c carbohydrate from Streptococcus mutans, which depended on the presence of the respective gacH homologs. Finally, nuclear magnetic resonance analysis of GAC confirmed that glycerol phosphate is attached to approximately 25% of the GAC N-acetylglucosamine side-chains at the C6 hydroxyl group. This previously unrecognized structural modification impacts host-pathogen interaction and has implications for vaccine design.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Glycerol / chemistry
Glycerol / metabolism*
Phosphates / chemistry
Phosphates / metabolism*
Polysaccharides, Bacterial / chemistry
Polysaccharides, Bacterial / metabolism*
Streptococcus / chemistry
Streptococcus / metabolism*

Substances

Phosphates
Polysaccharides, Bacterial
Glycerol

Abstract

Publication types

MeSH terms

Substances

Grants and funding