Recombinant fungal lectin as a new tool to investigate O-GlcNAcylation processes

Machon, Oriane; Baldini, Steffi F.; Ribeiro, João P; Steenackers, Agata; Varrot, Annabelle; Lefebvre, Tony; Imberty, Anne

Document type :

Article dans une revue scientifique

DOI :

10.1093/glycob/cww105

Permalink :

http://hdl.handle.net/20.500.12210/18663

Title :

Recombinant fungal lectin as a new tool to investigate O-GlcNAcylation processes

Author(s) :

Machon, Oriane [Auteur]
Centre de Recherches sur les Macromolécules Végétales [CERMAV]
Baldini, Steffi F. [Auteur]
Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF]
Ribeiro, João P [Auteur]
Centre de Recherches sur les Macromolécules Végétales [CERMAV]
Steenackers, Agata [Auteur]
Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF]
Varrot, Annabelle [Auteur]
Centre de Recherches sur les Macromolécules Végétales [CERMAV]
Lefebvre, Tony [Auteur]

Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF]
Unité de Glycobiologie Structurale et Fonctionnelle (UGSF) - UMR 8576
Imberty, Anne [Auteur]
Centre de Recherches sur les Macromolécules Végétales [CERMAV]

Journal title :

Glycobiology

Volume number :

Pages :

123-128

Publication date :

2017-01

ISSN :

0959-6658

English keyword(s) :

Fungal lectin
O-GlcNAcylation
rPVL

HAL domain(s) :

Chimie/Chimie théorique et/ou physique

English abstract : [en]

Glycosylation is a group of post-translational modifications that displays a large variety of structures and are implicated in a plethora of biological processes. Therefore, studying glycosylation requires different technical ...
Show more >Glycosylation is a group of post-translational modifications that displays a large variety of structures and are implicated in a plethora of biological processes. Therefore, studying glycosylation requires different technical approaches and reliable tools, lectins being part of them. Here, we describe the use of the recombinant mushroom lectin PVL to discriminate O-GlcNAcylation, a modification consisting in the attachment of a single N-acetylglucosamine residue to proteins confined within the cytosolic, nuclear and mitochondrial compartments. Recombinant PVL (Psathyrella velutina lectin) (rPVL) displays significantly stronger affinity for GlcNAc over Neu5Ac residues as verified by thermal shift assays and surface plasmon resonance experiments, being therefore an excellent alternative to WGA (wheat germ agglutinin). Labeling of rPVL with biotin or HRP (horseradish peroxidase) allows its useful and efficient utilization by western blot. The staining of whole cell lysates with labeled-rPVL was dramatically decreased in response to O-GlcNAc transferase knockdown and seen to increase after pharmacological blockade of O-GlcNAcase. Also, HRP-rPVL seemed to be more sensitive than the anti-O-GlcNAc antibody RL2. Thus, rPVL is a potent new tool to selectively detect O-GlcNAcylated proteins.Show less >

Language :

Anglais

Audience :

Non spécifiée

ANR Project :

Grenoble, une chimie bio-motivée

Administrative institution(s) :

CNRS
Université de Lille

Collections :

Unité de Glycobiologie Structurale et Fonctionnelle (UGSF) - UMR 8576

Research team(s) :

O-GlcNAcylation, signalisation cellulaire et cycle cellulaire

Submission date :

2020-02-12T15:45:16Z
2021-05-05T08:43:12Z

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2	20.500.12210/18663*	2021-05-05T08:37:16Z
1	20.500.12210/18663.1	2020-02-12T15:45:16Z

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Recombinant fungal lectin as a new tool ...

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