A glycoproteomic approach reveals that the S-layer glycoprotein of <i>Lactobacillus kefiri</i> CIDCA 83111 is <i>O</i>- and <i>N</i>-glycosylated

In Gram-positive bacteria, such as lactic acid bacteria, general glycosylation systems have not been documented so far. The aim of this work was to characterize in detail the glycosylation of the S-layer protein of <i>Lactobacillus kefiri</i> CIDCA 83111. A reductive β-elimination treatm...

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Autores principales: Cavallero, Gustavo J., Malamud, Mariano, Casabuono, Adriana C., Serradell, María de los Ángeles, Couto, Alicia S.
Formato: Articulo Preprint
Lenguaje:Inglés
Publicado: 2017
Materias:
Ciencias Exactas
Glycoproteomics
L. kefiri
S-layer glycoprotein
N-glycosylation
O-glycosylation
mass spectrometry
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/107301
https://www.sciencedirect.com/science/article/abs/pii/S1874391917301276
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-107301
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Ciencias Exactas
Glycoproteomics
L. kefiri
S-layer glycoprotein
N-glycosylation
O-glycosylation
mass spectrometry
spellingShingle Ciencias Exactas
Glycoproteomics
L. kefiri
S-layer glycoprotein
N-glycosylation
O-glycosylation
mass spectrometry
Cavallero, Gustavo J.
Malamud, Mariano
Casabuono, Adriana C.
Serradell, María de los Ángeles
Couto, Alicia S.
A glycoproteomic approach reveals that the S-layer glycoprotein of <i>Lactobacillus kefiri</i> CIDCA 83111 is <i>O</i>- and <i>N</i>-glycosylated
topic_facet Ciencias Exactas
Glycoproteomics
L. kefiri
S-layer glycoprotein
N-glycosylation
O-glycosylation
mass spectrometry
description In Gram-positive bacteria, such as lactic acid bacteria, general glycosylation systems have not been documented so far. The aim of this work was to characterize in detail the glycosylation of the S-layer protein of <i>Lactobacillus kefiri</i> CIDCA 83111. A reductive β-elimination treatment followed by anion exchange high performance liquid chromatography analysis was useful to characterize the <i>O</i>-glycosidic structures. MALDI-TOF mass spectrometry analysis confirmed the presence of oligosaccharides bearing from 5 to 8 glucose units carrying galacturonic acid. Further nanoHPLC-ESI analysis of the glycopeptides showed two <i>O</i>-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in <i>L. buchneri</i>, substituted on average with eight glucose residues and decorated with galacturonic acid and another <i>O</i>-glycosylated site on peptide 471–476, with a Glc<sub>5–8</sub>GalA<sub>2</sub> structure. As ten characteristic sequons (Asn-X-Ser/Thr) are present in the S-layer amino acid sequence, we performed a PNGase F digestion to release N-linked oligosaccharides. Anion exchange chromatography analysis showed mainly short N-linked chains. NanoHPLC-ESI in the positive and negative ion modes were useful to determine two different peptides substituted with short <i>N</i>-glycan structures. To our knowledge, this is the first description of the structure of <i>N</i>-glycans in S-layer glycoproteins from <i>Lactobacillus</i> species.
format Articulo
Preprint
author Cavallero, Gustavo J.
Malamud, Mariano
Casabuono, Adriana C.
Serradell, María de los Ángeles
Couto, Alicia S.
author_facet Cavallero, Gustavo J.
Malamud, Mariano
Casabuono, Adriana C.
Serradell, María de los Ángeles
Couto, Alicia S.
author_sort Cavallero, Gustavo J.
title A glycoproteomic approach reveals that the S-layer glycoprotein of <i>Lactobacillus kefiri</i> CIDCA 83111 is <i>O</i>- and <i>N</i>-glycosylated
title_short A glycoproteomic approach reveals that the S-layer glycoprotein of <i>Lactobacillus kefiri</i> CIDCA 83111 is <i>O</i>- and <i>N</i>-glycosylated
title_full A glycoproteomic approach reveals that the S-layer glycoprotein of <i>Lactobacillus kefiri</i> CIDCA 83111 is <i>O</i>- and <i>N</i>-glycosylated
title_fullStr A glycoproteomic approach reveals that the S-layer glycoprotein of <i>Lactobacillus kefiri</i> CIDCA 83111 is <i>O</i>- and <i>N</i>-glycosylated
title_full_unstemmed A glycoproteomic approach reveals that the S-layer glycoprotein of <i>Lactobacillus kefiri</i> CIDCA 83111 is <i>O</i>- and <i>N</i>-glycosylated
title_sort glycoproteomic approach reveals that the s-layer glycoprotein of <i>lactobacillus kefiri</i> cidca 83111 is <i>o</i>- and <i>n</i>-glycosylated
publishDate 2017
url http://sedici.unlp.edu.ar/handle/10915/107301
https://www.sciencedirect.com/science/article/abs/pii/S1874391917301276
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