Reference : Ligand Binding Study of Human PEBP1/RKIP: Interaction with Nucleotides and Raf-1 Peptide...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
http://hdl.handle.net/2268/124849
Ligand Binding Study of Human PEBP1/RKIP: Interaction with Nucleotides and Raf-1 Peptides Evidenced by NMR and Mass Spectrometry
English
Tavel, Laurette mailto [Université de Liège - ULg > Département de chimie (sciences) > Chimie biologique structurale >]
Jaquillard, Lucie mailto [Centre National de la Recherche Scientifique - CNRS > CBM-Orleans > > >]
Karsisiotis, Andreas mailto [Centre National de la Recherche Scientifique - CNRS > CBM-Orléans > > >]
Saab, Fabienne mailto [Centre National de la Recherche Scientifique - CNRS > CBM-Orleans > > >]
Jouvensal, Laurence mailto [Centre National de la Recherche Scientifique - CNRS > CBM-Orleans > > >]
Brans, Alain mailto [Université de Liège - ULg > > Centre d'ingénierie des protéines >]
Delmas, Agnès mailto [Centre National de la Recherche Scientifique - CNRS > CBM-Orleans > > >]
Schoentgen, Françoise mailto [Centre National de la Recherche Scientifique - CNRS > CBM-Orleans > > >]
Cadene, Martine mailto [Centre National de la Recherche Scientifique - CNRS > CBM-Orleans > > >]
Damblon, Christian mailto [Université de Liège - ULg > Département de chimie (sciences) > Chimie biologique structurale >]
27-Apr-2012
PLoS ONE
Public Library of Science
7(4): e36187
Yes (verified by ORBi)
International
International
1932-6203
San Franscisco
CA
[en] NMR ; RKIP ; protein-ligand interaction
[en] Background
Human Phosphatidylethanolamine binding protein 1 (hPEBP1) also known as Raf kinase inhibitory protein (RKIP), affects various cellular processes, and is implicated in metastasis formation and Alzheimer's disease. Human PEBP1 has also been shown to inhibit the Raf/MEK/ERK pathway. Numerous reports concern various mammalian PEBP1 binding ligands. However, since PEBP1 proteins from many different species were investigated, drawing general conclusions regarding human PEBP1 binding properties is rather difficult. Moreover, the binding site of Raf-1 on hPEBP1 is still unknown.
Methods/Findings
In the present study, we investigated human PEBP1 by NMR to determine the binding site of four different ligands: GTP, FMN, and one Raf-1 peptide in tri-phosphorylated and non-phosphorylated forms. The study was carried out by NMR in near physiological conditions, allowing for the identification of the binding site and the determination of the affinity constants KD for different ligands. Native mass spectrometry was used as an alternative method for measuring KD values.
Conclusions/Significance
Our study demonstrates and/or confirms the binding of hPEBP1 to the four studied ligands. All of them bind to the same region centered on the conserved ligand-binding pocket of hPEBP1. Although the affinities for GTP and FMN decrease as pH, salt concentration and temperature increase from pH 6.5/NaCl 0 mM/20°C to pH 7.5/NaCl 100 mM/30°C, both ligands clearly do bind under conditions similar to what is found in cells regarding pH, salt concentration and temperature. In addition, our work confirms that residues in the vicinity of the pocket rather than those within the pocket seem to be required for interaction with Raf-1.
Laboratoire de Chimie Biologique Structurale, ULg, Belgium ; Centre de Biophysique Moléculaire, Orléans, France ; Centre d'Ingénierie des Protéines - CIP
ANR (Agence Nationale pour la Recherche; grant ANR-08-BLAN-0033, France ; La Ligue contre le cancer, France ; Cancéropôle grand ouest AO2007-2010, France ; CNRS (Centre National pour la Recherche Scientifique), Régions Bretagne, Centre, Pays de la Loire et Poitou-Charentes (AO2007-2010), France ; Patrimoine de l′ULg ; EU-NMR program (RII3-026145)
METASUP
Researchers
http://hdl.handle.net/2268/124849
10.1371/journal.pone.0036187
http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0036187
Copyright: © 2012 Tavel et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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