Serine-rich repeat protein adhesins from Lactobacillus reuteri display strain specific glycosylation profiles

Dimitrios Latousakis; Ridvan Nepravishta; Martin Rejzek; Udo Wegmann; Gwenaelle Le Gall; Devon Kavanaugh; Ian Colquhoun; Steven Frese; Donald MacKenzie; Jens Walter; Jesus Angulo; Rob Field; Nathalie Juge

doi:10.1093/glycob/cwy100

Serine-rich repeat protein adhesins from Lactobacillus reuteri display strain specific glycosylation profiles

Dimitrios Latousakis
, Ridvan Nepravishta
, Martin Rejzek
, Udo Wegmann
, Gwenaelle Le Gall
, Devon Kavanaugh
, Ian Colquhoun
, Steven Frese
, Donald MacKenzie
, Jens Walter
, Jesus Angulo
, Rob Field
, Nathalie Juge (Lead Author)

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

21 Citations (Scopus)

42 Downloads (Pure)

Abstract

Lactobacillus reuteri is a gut symbiont inhabiting the gastrointestinal tract of numerous vertebrates. The surface-exposed Serine-Rich Repeat Protein (SRRP) is a major adhesin in Gram-positive bacteria. Using lectin and sugar nucleotide profiling of wild-type or L. reuteri isogenic mutants, MALDI-ToF-MS, LC-MS and GC-MS analyses of SRRPs, we showed that L. reuteri strains 100-23C (from rodent) and ATCC 53608 (from pig) can perform protein O-glycosylation and modify SRRP100-23 and SRRP53608 with Hex-Glc-GlcNAc and di-GlcNAc moieties, respectively. Furthermore, in vivo glycoengineering in E. coli led to glycosylation of SRRP53608 variants with α-GlcNAc and GlcNAcβ(1→6)GlcNAcα moieties. The glycosyltransferases involved in the modification of these adhesins were identified within the SecA2/Y2 accessory secretion system and their sugar nucleotide preference determined by saturation transfer difference NMR spectroscopy and differential scanning fluorimetry. Together, these findings provide novel insights into the cellular O-protein glycosylation pathways of gut commensal bacteria and potential routes for glycoengineering applications.

Original language	English
Pages (from-to)	45-58
Number of pages	14
Journal	Glycobiology
Volume	29
Issue number	1
Early online date	27 Oct 2018
DOIs	https://doi.org/10.1093/glycob/cwy100 https://doi.org/10.1093/glycob/cwy100
Publication status	Published - 1 Jan 2019

Keywords

Adhesins, Bacterial/chemistry
Glycosylation
Lactobacillus reuteri/chemistry
Mutation
Nuclear Magnetic Resonance, Biomolecular
Repetitive Sequences, Amino Acid

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10.1093/glycob/cwy100Licence: CC BY
10.1093/glycob/cwy100

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This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
Accepted author manuscript, 390 KBLicence: CC BY
Glycobiology_2019_Latousakis_etalFinal published version, 945 KBLicence: CC BY

Cite this

Latousakis, D., Nepravishta, R., Rejzek, M., Wegmann, U., Le Gall, G., Kavanaugh, D., Colquhoun, I., Frese, S., MacKenzie, D., Walter, J., Angulo, J., Field, R., & Juge, N. (2019). Serine-rich repeat protein adhesins from Lactobacillus reuteri display strain specific glycosylation profiles. Glycobiology, 29(1), 45-58. https://doi.org/10.1093/glycob/cwy100, https://doi.org/10.1093/glycob/cwy100

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title = "Serine-rich repeat protein adhesins from Lactobacillus reuteri display strain specific glycosylation profiles",

abstract = "Lactobacillus reuteri is a gut symbiont inhabiting the gastrointestinal tract of numerous vertebrates. The surface-exposed Serine-Rich Repeat Protein (SRRP) is a major adhesin in Gram-positive bacteria. Using lectin and sugar nucleotide profiling of wild-type or L. reuteri isogenic mutants, MALDI-ToF-MS, LC-MS and GC-MS analyses of SRRPs, we showed that L. reuteri strains 100-23C (from rodent) and ATCC 53608 (from pig) can perform protein O-glycosylation and modify SRRP100-23 and SRRP53608 with Hex-Glc-GlcNAc and di-GlcNAc moieties, respectively. Furthermore, in vivo glycoengineering in E. coli led to glycosylation of SRRP53608 variants with α-GlcNAc and GlcNAcβ(1→6)GlcNAcα moieties. The glycosyltransferases involved in the modification of these adhesins were identified within the SecA2/Y2 accessory secretion system and their sugar nucleotide preference determined by saturation transfer difference NMR spectroscopy and differential scanning fluorimetry. Together, these findings provide novel insights into the cellular O-protein glycosylation pathways of gut commensal bacteria and potential routes for glycoengineering applications.",

keywords = "Adhesins, Bacterial/chemistry, Glycosylation, Lactobacillus reuteri/chemistry, Mutation, Nuclear Magnetic Resonance, Biomolecular, Repetitive Sequences, Amino Acid",

author = "Dimitrios Latousakis and Ridvan Nepravishta and Martin Rejzek and Udo Wegmann and \{Le Gall\}, Gwenaelle and Devon Kavanaugh and Ian Colquhoun and Steven Frese and Donald MacKenzie and Jens Walter and Jesus Angulo and Rob Field and Nathalie Juge",

year = "2019",

month = jan,

day = "1",

doi = "10.1093/glycob/cwy100",

language = "English",

volume = "29",

pages = "45--58",

journal = "Glycobiology",

issn = "0959-6658",

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Latousakis, D, Nepravishta, R, Rejzek, M, Wegmann, U, Le Gall, G, Kavanaugh, D, Colquhoun, I, Frese, S, MacKenzie, D, Walter, J, Angulo, J, Field, R & Juge, N 2019, 'Serine-rich repeat protein adhesins from Lactobacillus reuteri display strain specific glycosylation profiles', Glycobiology, vol. 29, no. 1, pp. 45-58. https://doi.org/10.1093/glycob/cwy100, https://doi.org/10.1093/glycob/cwy100

TY - JOUR

T1 - Serine-rich repeat protein adhesins from Lactobacillus reuteri display strain specific glycosylation profiles

AU - Latousakis, Dimitrios

AU - Nepravishta, Ridvan

AU - Rejzek, Martin

AU - Wegmann, Udo

AU - Le Gall, Gwenaelle

AU - Kavanaugh, Devon

AU - Colquhoun, Ian

AU - Frese, Steven

AU - MacKenzie, Donald

AU - Walter, Jens

AU - Angulo, Jesus

AU - Field, Rob

AU - Juge, Nathalie

PY - 2019/1/1

Y1 - 2019/1/1

N2 - Lactobacillus reuteri is a gut symbiont inhabiting the gastrointestinal tract of numerous vertebrates. The surface-exposed Serine-Rich Repeat Protein (SRRP) is a major adhesin in Gram-positive bacteria. Using lectin and sugar nucleotide profiling of wild-type or L. reuteri isogenic mutants, MALDI-ToF-MS, LC-MS and GC-MS analyses of SRRPs, we showed that L. reuteri strains 100-23C (from rodent) and ATCC 53608 (from pig) can perform protein O-glycosylation and modify SRRP100-23 and SRRP53608 with Hex-Glc-GlcNAc and di-GlcNAc moieties, respectively. Furthermore, in vivo glycoengineering in E. coli led to glycosylation of SRRP53608 variants with α-GlcNAc and GlcNAcβ(1→6)GlcNAcα moieties. The glycosyltransferases involved in the modification of these adhesins were identified within the SecA2/Y2 accessory secretion system and their sugar nucleotide preference determined by saturation transfer difference NMR spectroscopy and differential scanning fluorimetry. Together, these findings provide novel insights into the cellular O-protein glycosylation pathways of gut commensal bacteria and potential routes for glycoengineering applications.

AB - Lactobacillus reuteri is a gut symbiont inhabiting the gastrointestinal tract of numerous vertebrates. The surface-exposed Serine-Rich Repeat Protein (SRRP) is a major adhesin in Gram-positive bacteria. Using lectin and sugar nucleotide profiling of wild-type or L. reuteri isogenic mutants, MALDI-ToF-MS, LC-MS and GC-MS analyses of SRRPs, we showed that L. reuteri strains 100-23C (from rodent) and ATCC 53608 (from pig) can perform protein O-glycosylation and modify SRRP100-23 and SRRP53608 with Hex-Glc-GlcNAc and di-GlcNAc moieties, respectively. Furthermore, in vivo glycoengineering in E. coli led to glycosylation of SRRP53608 variants with α-GlcNAc and GlcNAcβ(1→6)GlcNAcα moieties. The glycosyltransferases involved in the modification of these adhesins were identified within the SecA2/Y2 accessory secretion system and their sugar nucleotide preference determined by saturation transfer difference NMR spectroscopy and differential scanning fluorimetry. Together, these findings provide novel insights into the cellular O-protein glycosylation pathways of gut commensal bacteria and potential routes for glycoengineering applications.

KW - Adhesins, Bacterial/chemistry

KW - Glycosylation

KW - Lactobacillus reuteri/chemistry

KW - Mutation

KW - Nuclear Magnetic Resonance, Biomolecular

KW - Repetitive Sequences, Amino Acid

UR - https://www.scopus.com/pages/publications/85058750909

U2 - 10.1093/glycob/cwy100

DO - 10.1093/glycob/cwy100

M3 - Article

C2 - 30371779

SN - 0959-6658

VL - 29

SP - 45

EP - 58

JO - Glycobiology

JF - Glycobiology

IS - 1

ER -

Serine-rich repeat protein adhesins from Lactobacillus reuteri display strain specific glycosylation profiles

Abstract

Keywords

Access to Document

Other files and links

Jesus Angulo

Gwenaelle Le Gall

Towards understanding the glycan code: next generation of structural glycobiology tools for accurate description of protein-glycan complexes.

Cite this

Serine-rich repeat protein adhesins from Lactobacillus reuteri display strain specific glycosylation profiles

Abstract

Keywords

Access to Document

Other files and links

Profiles

Jesus Angulo

Gwenaelle Le Gall

Projects

Towards understanding the glycan code: next generation of structural glycobiology tools for accurate description of protein-glycan complexes.

Cite this