Ligand binding study of human PEBP1/RKIP: interaction with nucleotides and Raf-1 peptides evidenced by NMR and mass spectrometry.

PloS One
Laurette TavelChristian Damblon

Abstract

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. 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 K(D) for different ligands. Native mass spectrometry was used as an alternative method for measuring K(D) values. 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. Altho...Continue Reading

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Citations

Nov 9, 2012·Cell Communication and Signaling : CCS·Tomonori KanekoShawn Sc Li
Jan 30, 2013·Journal of Cellular Physiology·Fahd Al-MullaKam C Yeung
Jul 13, 2014·Tumour Biology : the Journal of the International Society for Oncodevelopmental Biology and Medicine·Bin LiJianhui Gu
Apr 6, 2017·Tumour Biology : the Journal of the International Society for Oncodevelopmental Biology and Medicine·SoHyun LeeBenjamin Bonavida
Jan 22, 2020·Cellular and Molecular Life Sciences : CMLS·Françoise Schoentgen, Slavica Jonic
Jun 2, 2016·Journal of the American Society for Mass Spectrometry·Guillaume GabantMartine Cadene

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Methods Mentioned

BETA
X-ray
NMR
surface plasmon resonance
ion exchange chromatography
chemical exchange
gel filtration

Software Mentioned

SigmaPlot

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