Immunostimulation by Lactobacillus kefiri S-layer proteins with distinct glycosylation patterns requires different lectin partners

S-layer (glyco)-proteins (SLPs) form a nanostructured envelope that covers the surface of different prokaryotes and show immunomodulatory activity. Previously, we have demonstrated that the S-layer glycoprotein from probiotic Lactobacillus kefiri CIDCA 8348 (SLP-8348) is recognized by Mincle (macrop...

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Autores principales: Malamud, Mariano, Cavallero, Gustavo J., Casabuono, Adriana C., Lepenies, Bernd, Serradell, María de los Ángeles, Couto, Alicia S.
Formato: Articulo
Lenguaje:Inglés
Publicado: 2020
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Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/124330
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id I19-R120-10915-124330
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Bioquímica
Lactobacillus kefiri
S-layer protein
Glycosylation
Innate immunity
Mass spectrometry (MS)
N-linked glycosylation
Gram-positive bacteria
C-type lectin receptors
O-linked glycosylation
spellingShingle Bioquímica
Lactobacillus kefiri
S-layer protein
Glycosylation
Innate immunity
Mass spectrometry (MS)
N-linked glycosylation
Gram-positive bacteria
C-type lectin receptors
O-linked glycosylation
Malamud, Mariano
Cavallero, Gustavo J.
Casabuono, Adriana C.
Lepenies, Bernd
Serradell, María de los Ángeles
Couto, Alicia S.
Immunostimulation by Lactobacillus kefiri S-layer proteins with distinct glycosylation patterns requires different lectin partners
topic_facet Bioquímica
Lactobacillus kefiri
S-layer protein
Glycosylation
Innate immunity
Mass spectrometry (MS)
N-linked glycosylation
Gram-positive bacteria
C-type lectin receptors
O-linked glycosylation
description S-layer (glyco)-proteins (SLPs) form a nanostructured envelope that covers the surface of different prokaryotes and show immunomodulatory activity. Previously, we have demonstrated that the S-layer glycoprotein from probiotic Lactobacillus kefiri CIDCA 8348 (SLP-8348) is recognized by Mincle (macrophage inducible C-type lectin receptor), and its adjuvanticity depends on the integrity of its glycans. However, the glycan's structure has not been described so far. Herein, we analyze the glycosylation pattern of three SLPs, SLP-8348, SLP-8321, and SLP-5818, and explore how these patterns impact their recognition by C-type lectin receptors and the immunomodulatory effect of the L. kefiri SLPs on antigen-presenting cells. High-performance anion-exchange chromatography–pulse amperometric detector performed after β-elimination showed glucose as the major component in the O-glycans of the three SLPs; however, some differences in the length of hexose chains were observed. No N-glycosylation signals were detected in SLP-8348 and SLP-8321, but SLP-5818 was observed to have two sites carrying complex N-glycans based on a site-specific analysis and a glycomic workflow of the permethylated glycans. SLP-8348 was previously shown to enhance LPS-induced activation on both RAW264.7 macrophages and murine bone marrow–derived dendritic cells; we now show that SLP-8321 and SLP-5818 have a similar effect regardless of the differences in their glycosylation patterns. Studies performed with bone marrow–derived dendritic cells from C-type lectin receptor–deficient mice revealed that the immunostimulatory activity of SLP-8321 depends on its recognition by Mincle, whereas SLP-5818's effects are dependent on SignR3 (murine ortholog of human DC-SIGN). These findings encourage further investigation of both the potential application of these SLPs as new adjuvants and the protein glycosylation mechanisms in these bacteria.
format Articulo
Articulo
author Malamud, Mariano
Cavallero, Gustavo J.
Casabuono, Adriana C.
Lepenies, Bernd
Serradell, María de los Ángeles
Couto, Alicia S.
author_facet Malamud, Mariano
Cavallero, Gustavo J.
Casabuono, Adriana C.
Lepenies, Bernd
Serradell, María de los Ángeles
Couto, Alicia S.
author_sort Malamud, Mariano
title Immunostimulation by Lactobacillus kefiri S-layer proteins with distinct glycosylation patterns requires different lectin partners
title_short Immunostimulation by Lactobacillus kefiri S-layer proteins with distinct glycosylation patterns requires different lectin partners
title_full Immunostimulation by Lactobacillus kefiri S-layer proteins with distinct glycosylation patterns requires different lectin partners
title_fullStr Immunostimulation by Lactobacillus kefiri S-layer proteins with distinct glycosylation patterns requires different lectin partners
title_full_unstemmed Immunostimulation by Lactobacillus kefiri S-layer proteins with distinct glycosylation patterns requires different lectin partners
title_sort immunostimulation by lactobacillus kefiri s-layer proteins with distinct glycosylation patterns requires different lectin partners
publishDate 2020
url http://sedici.unlp.edu.ar/handle/10915/124330
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