Selective Exoenzymatic Labeling of Lipooligosaccharides of Neisseria gonorrhoeae with α2,6-Sialoside Analogues

Hanna de Jong; Maria J. Moure; Jet E. M. Hartman; Gerlof P. Bosman; Jun Yang Ong; Bart W. Bardoel; Geert-Jan Boons; Marc M. S. M. Wosten; Tom Wennekes

doi:10.1002/cbic.202200340

Selective Exoenzymatic Labeling of Lipooligosaccharides of Neisseria gonorrhoeae with α2,6-Sialoside Analogues

Hanna de Jong
, Maria J. Moure
, Jet E. M. Hartman
, Gerlof P. Bosman
, Jun Yang Ong
, Bart W. Bardoel
, Geert-Jan Boons
, Marc M. S. M. Wosten
, Tom Wennekes

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

Abstract

The interactions between bacteria and their host often rely on recognition processes that involve host or bacterial glycans. Glycoengineering techniques make it possible to modify and study the glycans on the host's eukaryotic cells, but only a few are available for the study of bacterial glycans. Here, we have adapted selective exoenzymatic labeling (SEEL), a chemical reporter strategy, to label the lipooligosaccharides of the bacterial pathogen Neisseria gonorrhoeae, using the recombinant glycosyltransferase ST6Gal1, and three synthetic CMP-sialic acid derivatives. We show that SEEL treatment does not affect cell viability and can introduce an α2,6-linked sialic acid with a reporter group on the lipooligosaccharides by Western blot, flow cytometry and fluorescent microscopy. This new bacterial glycoengineering technique allows for the precise modification, here with α2,6-sialoside derivatives, and direct detection of specific surface glycans on live bacteria, which will aid in further unravelling the precise biological functions of bacterial glycans.

Original language	English
Article number	e202200340
Number of pages	9
Journal	ChemBioChem
Volume	23
Issue number	19
Early online date	23 Aug 2022
DOIs	https://doi.org/10.1002/cbic.202200340
Publication status	Published - 6 Oct 2022

Bibliographical note

Funding Information:
We thank Jos van Putten for generously providing the strains described in this research. We thank Esther van ‘t Veld and Richard Wubbolts of the Centre for Cell Imaging (CCI) of the faculty of Veterinary Medicine for technical support regarding imaging. Research in this publication was supported by the Netherlands Foundation for Scientific Research (NWO) via a VIDI grant (723.014.005 to TW) and BBOL grant (737.016.013). JYO and TW received funding from the European Union's Horizon 2020 Marie Skłodowska‐Curie Actions for the Innovative Training Network “Sweet Crosstalk” under the grant agreement No 814102. Neisseria gonorrhoeae

Funding Information:
We thank Jos van Putten for generously providing the Neisseria gonorrhoeae strains described in this research. We thank Esther van ‘t Veld and Richard Wubbolts of the Centre for Cell Imaging (CCI) of the faculty of Veterinary Medicine for technical support regarding imaging. Research in this publication was supported by the Netherlands Foundation for Scientific Research (NWO) via a VIDI grant (723.014.005 to TW) and BBOL grant (737.016.013). JYO and TW received funding from the European Union's Horizon 2020 Marie Skłodowska-Curie Actions for the Innovative Training Network “Sweet Crosstalk” under the grant agreement No 814102.

Publisher Copyright:
© 2022 The Authors. ChemBioChem published by Wiley-VCH GmbH.

Keywords

glycobiology
glycoengineering
lipooligosaccharides
sialic acid

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ChemBioChem - 2022 - Jong - Selective Exoenzymatic Labeling of Lipooligosaccharides of Neisseria gonorrhoeae with 2Final published version, 1.23 MBLicence: CC BY

Cite this

@article{9f9558a1afff49c682329af6faf09b21,

title = "Selective Exoenzymatic Labeling of Lipooligosaccharides of Neisseria gonorrhoeae with α2,6-Sialoside Analogues",

abstract = "The interactions between bacteria and their host often rely on recognition processes that involve host or bacterial glycans. Glycoengineering techniques make it possible to modify and study the glycans on the host's eukaryotic cells, but only a few are available for the study of bacterial glycans. Here, we have adapted selective exoenzymatic labeling (SEEL), a chemical reporter strategy, to label the lipooligosaccharides of the bacterial pathogen Neisseria gonorrhoeae, using the recombinant glycosyltransferase ST6Gal1, and three synthetic CMP-sialic acid derivatives. We show that SEEL treatment does not affect cell viability and can introduce an α2,6-linked sialic acid with a reporter group on the lipooligosaccharides by Western blot, flow cytometry and fluorescent microscopy. This new bacterial glycoengineering technique allows for the precise modification, here with α2,6-sialoside derivatives, and direct detection of specific surface glycans on live bacteria, which will aid in further unravelling the precise biological functions of bacterial glycans.",

keywords = "glycobiology, glycoengineering, lipooligosaccharides, sialic acid",

author = "\{de Jong\}, Hanna and Moure, \{Maria J.\} and Hartman, \{Jet E. M.\} and Bosman, \{Gerlof P.\} and Ong, \{Jun Yang\} and Bardoel, \{Bart W.\} and Geert-Jan Boons and Wosten, \{Marc M. S. M.\} and Tom Wennekes",

note = "Funding Information: We thank Jos van Putten for generously providing the strains described in this research. We thank Esther van {\textquoteleft}t Veld and Richard Wubbolts of the Centre for Cell Imaging (CCI) of the faculty of Veterinary Medicine for technical support regarding imaging. Research in this publication was supported by the Netherlands Foundation for Scientific Research (NWO) via a VIDI grant (723.014.005 to TW) and BBOL grant (737.016.013). JYO and TW received funding from the European Union's Horizon 2020 Marie Sk{\l}odowska‐Curie Actions for the Innovative Training Network “Sweet Crosstalk” under the grant agreement No 814102. Neisseria gonorrhoeae Funding Information: We thank Jos van Putten for generously providing the Neisseria gonorrhoeae strains described in this research. We thank Esther van {\textquoteleft}t Veld and Richard Wubbolts of the Centre for Cell Imaging (CCI) of the faculty of Veterinary Medicine for technical support regarding imaging. Research in this publication was supported by the Netherlands Foundation for Scientific Research (NWO) via a VIDI grant (723.014.005 to TW) and BBOL grant (737.016.013). JYO and TW received funding from the European Union's Horizon 2020 Marie Sk{\l}odowska-Curie Actions for the Innovative Training Network “Sweet Crosstalk” under the grant agreement No 814102. Publisher Copyright: {\textcopyright} 2022 The Authors. ChemBioChem published by Wiley-VCH GmbH.",

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doi = "10.1002/cbic.202200340",

language = "English",

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AU - de Jong, Hanna

AU - Moure, Maria J.

AU - Hartman, Jet E. M.

AU - Bosman, Gerlof P.

AU - Ong, Jun Yang

AU - Bardoel, Bart W.

AU - Boons, Geert-Jan

AU - Wosten, Marc M. S. M.

AU - Wennekes, Tom

N1 - Funding Information: We thank Jos van Putten for generously providing the strains described in this research. We thank Esther van ‘t Veld and Richard Wubbolts of the Centre for Cell Imaging (CCI) of the faculty of Veterinary Medicine for technical support regarding imaging. Research in this publication was supported by the Netherlands Foundation for Scientific Research (NWO) via a VIDI grant (723.014.005 to TW) and BBOL grant (737.016.013). JYO and TW received funding from the European Union's Horizon 2020 Marie Skłodowska‐Curie Actions for the Innovative Training Network “Sweet Crosstalk” under the grant agreement No 814102. Neisseria gonorrhoeae Funding Information: We thank Jos van Putten for generously providing the Neisseria gonorrhoeae strains described in this research. We thank Esther van ‘t Veld and Richard Wubbolts of the Centre for Cell Imaging (CCI) of the faculty of Veterinary Medicine for technical support regarding imaging. Research in this publication was supported by the Netherlands Foundation for Scientific Research (NWO) via a VIDI grant (723.014.005 to TW) and BBOL grant (737.016.013). JYO and TW received funding from the European Union's Horizon 2020 Marie Skłodowska-Curie Actions for the Innovative Training Network “Sweet Crosstalk” under the grant agreement No 814102. Publisher Copyright: © 2022 The Authors. ChemBioChem published by Wiley-VCH GmbH.

PY - 2022/10/6

Y1 - 2022/10/6

N2 - The interactions between bacteria and their host often rely on recognition processes that involve host or bacterial glycans. Glycoengineering techniques make it possible to modify and study the glycans on the host's eukaryotic cells, but only a few are available for the study of bacterial glycans. Here, we have adapted selective exoenzymatic labeling (SEEL), a chemical reporter strategy, to label the lipooligosaccharides of the bacterial pathogen Neisseria gonorrhoeae, using the recombinant glycosyltransferase ST6Gal1, and three synthetic CMP-sialic acid derivatives. We show that SEEL treatment does not affect cell viability and can introduce an α2,6-linked sialic acid with a reporter group on the lipooligosaccharides by Western blot, flow cytometry and fluorescent microscopy. This new bacterial glycoengineering technique allows for the precise modification, here with α2,6-sialoside derivatives, and direct detection of specific surface glycans on live bacteria, which will aid in further unravelling the precise biological functions of bacterial glycans.

AB - The interactions between bacteria and their host often rely on recognition processes that involve host or bacterial glycans. Glycoengineering techniques make it possible to modify and study the glycans on the host's eukaryotic cells, but only a few are available for the study of bacterial glycans. Here, we have adapted selective exoenzymatic labeling (SEEL), a chemical reporter strategy, to label the lipooligosaccharides of the bacterial pathogen Neisseria gonorrhoeae, using the recombinant glycosyltransferase ST6Gal1, and three synthetic CMP-sialic acid derivatives. We show that SEEL treatment does not affect cell viability and can introduce an α2,6-linked sialic acid with a reporter group on the lipooligosaccharides by Western blot, flow cytometry and fluorescent microscopy. This new bacterial glycoengineering technique allows for the precise modification, here with α2,6-sialoside derivatives, and direct detection of specific surface glycans on live bacteria, which will aid in further unravelling the precise biological functions of bacterial glycans.

KW - glycobiology

KW - glycoengineering

KW - lipooligosaccharides

KW - sialic acid

UR - https://www.scopus.com/pages/publications/85136454028

U2 - 10.1002/cbic.202200340

DO - 10.1002/cbic.202200340

M3 - Article

SN - 1439-4227

VL - 23

JO - ChemBioChem

JF - ChemBioChem

IS - 19

M1 - e202200340

ER -

Selective Exoenzymatic Labeling of Lipooligosaccharides of Neisseria gonorrhoeae with α2,6-Sialoside Analogues

Abstract

Bibliographical note

Keywords

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