IndraLab
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"In addition to confirming that locostatin prevents association of RKIP with Raf-1 under conditions where RKIP is preincubated with locostatin, we demonstrate in this report that locostatin added at the same time as RKIP to a Raf-1 sample results in disruption of binding of RKIP to Raf-1 when the sample is incubated at 37degreesC for 6 h, but not at 4degreesC for 30 min (XREF_FIG)."
sparser
"While there is evidence that phosphorylation of Ser388, Ser389, and Tyr341 on Raf-1 increases affinity of Raf-1 for RKIP (Ref. xref and R. Leo Brady, personal communication), there are also results to suggest that interaction of Raf-1 with RKIP instead blocks the phosphorylation of these same residues on Raf-1. xref The ligand-binding pocket could also conceivably accommodate Tyr and Asp residues, though with lower expected affinity than pTyr. xref It is possible that phosphorylation of Tyr residues on Raf-1 such as its Tyr341 residue (Ref. xref and R. Leo Brady, personal communication) and GRK2 could modulate the affinity of binding of these proteins to RKIP."
sparser
"We then performed double immunofluorescence in cultured neurons at 6 h after OGD/R and found that, compared with control group, there was a significant decrease in the co-location between PEBP1 and Raf-1, which means a weakening of the interaction between PEBP1 and Raf-1 (Figure xref )."
sparser
"Furthermore, analysis of docked complexes from the above servers showed the occurrence of six C-Raf (Tyr340, Tyr341, Trp342, Glu345, Arg398, and Lys399) and eight RKIP (Asp70, Ala73, Lys80, Tyr81, His86, Gly110, Tyr181, and Glu182) actively interacting amino acid residues observed as common in docked complexes from both servers ( xref )."
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"Ideally, as shown in Figures XREF_FIG - XREF_FIG and Supplementary Figure XREF_SUPPLEMENTARY, under I/R condition, the phosphorylation and the binding to Raf-1 of PEBP1 and GFP-PEBP1 shared the same trend, suggesting that fusion with GFP did not induce unwanted side effects of PEBP1 in this study."
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"reported that cerebral ischemia induced association between RKIP and Raf-1 resulted in the inhibition of the ERK signaling cascade through an inhibition of Src mediated Raf-1 phosphorylation at Tyr340/341 residues, suggesting that RKIP is involved in biphasic phosphorylation of ERK after brain ischemia."
sparser
"To see whether the expression of PEBP1 causes silencing of HIV by inactivating NF‐κB signaling pathway and whether this is through its interacting with Raf1 and IKK, we performed a co‐immunoprecipitation (Co‐IP) assay and revealed that PEBP1 not only interacted with Raf1 but also with IKK in C11 HIV latency cell model (Fig xref B)."
sparser
"In addition to confirming that locostatin prevents association of RKIP with Raf-1 under conditions where RKIP is preincubated with locostatin ( xref and Ref. xref ), we demonstrate in this report that locostatin added at the same time as RKIP to a Raf-1 sample results in disruption of binding of RKIP to Raf-1 when the sample is incubated at 37°C for 6 h, but not at 4°C for 30 min ( xref )."
sparser
"Another report suggested that locostatin does not disrupt the interaction of RKIP with Raf-1; however, in that study, locostatin, RKIP and Raf-1 were mixed together at the same time and the subsequent incubation was done at 4°C for only 30 min. xref These conditions are not sufficient to observe disruption of the RKIP-Raf-1 interaction ( xref )."
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"Here, using a combination of mutational approaches, biochemical studies, peptide arrays and plasmon surface resonance (BIAcore), we fine map and characterize a minimal 24 amino acid long RKIP binding domain in the Raf-1 N-region, which consists of constitutive elements at both flanks and a center element that is regulated by phosphorylation and enhances the re-binding of RKIP to Raf-1 in the later phase of mitogen stimulation."
sparser
"Locostatin is an inhibitor of cell migration xref and cell-substratum adhesion xref that covalently binds Raf kinase inhibitor protein (RKIP). xref Locostatin disrupts the interaction of RKIP with Raf-1 kinase. xref RKIP has a number of functions, including modulation of the activity of Raf-1 kinase, xref , xref G protein-coupled receptor kinase 2 (GRK2), xref and proteins involved in nuclear factor κB activation, including inhibitor of κB kinase α (IKKα) xref , xref and transforming growth factor β-activated kinase 1 (TAK1). xref , xref In addition, RKIP binds a number of endogenous small-molecule ligands, most notably phosphatidylethanolamine, and therefore is also known as phosphatidylethanolamine-binding protein (PEBP). xref , xref There is evidence that RKIP/PEBP can bind morphine, xref , xref morphine glucuronides, xref , xref nucleotides, xref and a pyrazolopyrimidine phosphodiesterase-5 inhibitor. xref RKIP has also been identified as a potential odorant-binding protein. xref However, the effects of these various small molecules on the function of RKIP are unclear and, at least in the case of phosphatidylethanoloamine and morphine glucuronides, these interactions are of low affinity and appear nonspecific. xref "
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"23 Although the mechanism of this interaction needs to be further defined, the interactions of 15HETE-PE with RKIP observed in human airway epithelial cells are consistent with a previous study which showed phospholipid binding in the RKIP binding pocket inhibited both Raf-1 binding to RKIP and its phosphorylation."
sparser
"From the above overall analysis, the mapped 13 interface binding sites acquired from the C-Raf/RKIP interaction can be considered as druggable binding sites (“hot spots”) for future designing of small molecule inhibitors that may inhibit the protein-protein interaction between C-Raf and RKIP."
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"As has been made clear in numerous other chapters, RKIP binds to Raf-1, one of the few known endogenous inhibitors of the mitogen activated protein kinase (MAPK) pathway and inhibits Raf-1-mediated activation of the mitogen activated protein kinase (MAPK) extracellular signal regulated kinase (ERK) by preventing phosphorylation of Raf-1."
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"Notably, compared with that in endogenous PEBP1, OGD/R exerted a similar induction of the phosphorylation of GFP-PEBP1 at Ser153 and inhibition of the interaction between GFP-PEBP1 and Raf-1, which was almost abolished by the mutation of Ser153 to Ala, therefore supporting Ser153 as a key site for OGD/R induced PEBP1 phosphorylation."