Arginine 469 is a pivotal residue for the Hsc70-GlcNAc-binding property.
The members of the 70kDa-heat shock proteins (HSP70) family play numerous fundamental functions in the cell such as promoting the assembly of multimeric complexes or helping the correct folding of nascent proteins to take place. In numerous previous studies we demonstrated that Hsp70 and its constitutive isoform Hsc70 are endowed of a GlcNAc-binding activity. The molecular modeling of the substrate binding domain of Hsc70 and in silico docking experiments using Ser/Thr-O-GlcNAc motifs allowed to define the potential carbohydrate-recognition region and to point out the crucial position of Arg469 as an amino-acid directly interacting with the sugar moiety. We cloned a flagged Hsc70 in a pCMV.SPORT6 vector and we showed that the mutation R469A decreased the GlcNAc-binding property of the chaperone of around 70%. This is the first work reporting the localization of the GlcNAc-binding domain of a member of the HSP70 family.
Guinez
Céline
C
CNRS-UMR 8576, Unit of Structural and Functional Glycobiology, IFR 147, Université de Lille 1, Cité Scientifique, 59655 Villeneuve d'Ascq, France.
Mir
Anne-Marie
AM
Martin
Nathalie
N
Leprince
Dominique
D
Michalski
Jean-Claude
JC
Vergoten
Gérard
G
Lefebvre
Tony
T
D000117:Acetylglucosamine / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D001120:Arginine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D019556:COS Cells; D002522:Chlorocebus aethiops; D050883:HSC70 Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D009154:Mutation; D011485:Protein Binding; D017434:Protein Structure, Tertiary
D050883:HSC70 Heat-Shock Proteins; C494371:HSPA8 protein, human; D001120:Arginine; D000117:Acetylglucosamine