A Novel Peptide Probe for Studying the Transbilayer Movement of Phosphatidylethanolamine

Aoki, Yoshiko; Uenaka, Toshimitsu; Aoki, Junken; Umeda, Masato; Inoue, Keizō

doi:10.1093/oxfordjournals.jbchem.a124522

Cited by 60 publications

(49 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Construction of FL-SA-Ro-Ro 09-0198, kindly provided by Roche Applied Science, was biotinylated using Biotin (Long Arm) NHS (Vector Laboratories) (14). FL-SA (Vector Laboratories) was mixed with biotinylated Ro, and the resulting FL-SA-Ro complexes were purified by gel filtration, as described (16).…”

Section: Quantitation Of Hpebp4 Expression By Real Time Fluorescencementioning

confidence: 99%

A Novel Human Phosphatidylethanolamine-binding Protein Resists Tumor Necrosis Factor α-induced Apoptosis by Inhibiting Mitogen-activated Protein Kinase Pathway Activation and Phosphatidylethanolamine Externalization

Wang

Liu

et al. 2004

Journal of Biological Chemistry

115

View full text Add to dashboard Cite

The phosphatidylethanolamine (PE)-binding proteins (PEBPs) are an evolutionarily conserved family of proteins with pivotal biological functions. Here we describe the cloning and functional characterization of a novel family member, human phosphatidylethanolaminebinding protein 4 (hPEBP4). hPEBP4 is expressed in most human tissues and highly expressed in tumor cells. Its expression in tumor cells is further enhanced upon tumor necrosis factor (TNF) ␣ treatment, whereas hPEBP4 normally co-localizes with lysosomes, TNF␣ stimulation triggers its transfer to the cell membrane, where it binds to Raf-1 and MEK1. L929 cells overexpressing hPEBP4 are resistant to both TNF␣-induced ERK1/2, MEK1, and JNK activation and TNF␣-mediated apoptosis. Co-precipitation and in vitro protein binding assay demonstrated that hPEBP4 interacts with Raf-1 and MEK1. A truncated form of hPEBP4, lacking the PE-binding domain, maintains lysosomal co-localization but has no effect on cellular responses to TNF␣. Given that MCF-7 breast cancer cells expressed hPEBP4 at a high level, small interfering RNA was used to silence the expression of hPEBP4. We demonstrated that down-regulation of hPEBP4 expression sensitizes MCF-7 breast cancer cells to TNF␣-induced apoptosis. hPEBP4 appears to promote cellular resistance to TNF-induced apoptosis by inhibiting activation of the Raf-1/MEK/ERK pathway, JNK, and PE externalization, and the conserved region of PE-binding domain appears to play a vital role in this biological activity of hPEBP4.The phosphatidylethanolamine-binding protein (PEBP) 1 family consists of a number of 21-23-kDa basic proteins, first identified in bovine brain, with preferential in vitro affinity for phosphatidylethanolamine, a component of the cell membrane. This family is an evolutionarily conserved group found in species of flowering plants (Antirrhinum (1)), parasites (Plasmodium falciparium (2)), nematodes (Toxocara canis (3)), insects (Drosophila melanogaster (4)), and mammals, including cattle, monkeys, and humans (5). A number of functions have been suggested for the mammalian PEBP proteins, including lipid binding and inhibition of serine proteases (6). These proteins can also act as precursors for a bioactive peptide HCNP (hippocampal cholinergic neurostimulating peptide), important in hippocampus development (5). Plant PEBP homologues are involved in the control of a morphogenic switch between shoot growth and flower structures (7). Yeast two-hybrid screen analysis has shown that human PEBP1 (hPEBP1, also called Raf kinase inhibitory protein or RKIP) acts as a suppressor of Raf-1 kinase activity and mitogen-activated protein kinase signaling in fibroblasts via its ability to sequester and inactivate Raf-1 and MEK1 (8, 9). Both Raf-1 and MEK bind to the highly conserved phosphatidylethanolamine-binding domain of hPEBP; hPEBP induces dissociation of Raf-1⅐MEK complexes and behaves as a competitive inhibitor of MEK phosphorylation. Mapping of the binding domains has shown that MEK and Raf-1 bind to overlapping sites in hPEBP...

show abstract

Section: Quantitation Of Hpebp4 Expression By Real Time Fluorescencementioning

confidence: 99%

A Novel Human Phosphatidylethanolamine-binding Protein Resists Tumor Necrosis Factor α-induced Apoptosis by Inhibiting Mitogen-activated Protein Kinase Pathway Activation and Phosphatidylethanolamine Externalization

Wang

Liu

et al. 2004

Journal of Biological Chemistry

115

View full text Add to dashboard Cite

show abstract

“…Therefore, we investigated the amount of surface-exposed PE in our pdr5⌬ and pdr5⌬ yor1⌬ cells using the tetracyclic peptide antibiotic Ro 09-0198 (Ro), which specifically binds to PE (Wakamatsu et al, 1990;Umeda and Emoto, 1999). Ro binding to PE results in cytolysis (Aoki et al, 1994;Kato et al, 2002), so the amount of surfaceexposed PE can easily be estimated by measuring the overall sensitivity of the cell to Ro. As shown in Figure 1A, treatment of wild-type cells with 50 M Ro caused a reduction in their growth rate (to ϳ30%).…”

Section: Accumulation Of Pe In the Outer Leaflet Of Pdr5⌬ And Pdr5⌬ Ymentioning

confidence: 99%

The Rim101 Pathway Is Involved in Rsb1 Expression Induced by Altered Lipid Asymmetry

Ikeda

Kihara

Denpoh

et al. 2008

MBoC

View full text Add to dashboard Cite

Biological membranes consist of lipid bilayers. The lipid compositions between the two leaflets of the plasma membrane differ, generating lipid asymmetry. Maintenance of proper lipid asymmetry is physiologically quite important, and its collapse induces several cellular responses including apoptosis and platelet coagulation. Thus, a change in lipid asymmetry must be restored to maintain "lipid asymmetry homeostasis." However, to date no lipid asymmetry-sensing proteins or any related downstream signaling pathways have been identified. We recently demonstrated that expression of the putative yeast sphingoid long-chain base transporter/translocase Rsb1 is induced when glycerophospholipid asymmetry is altered. Using mutant screening, we determined that the pH-responsive Rim101 pathway, the protein kinase Mck1, and the transcription factor Mot3 all act in lipid asymmetry signaling, and that the Rim101 pathway was activated in response to a change in lipid asymmetry. The activated transcription factor Rim101 induces Rsb1 expression via repression of another transcription repressor, Nrg1. Changes in lipid asymmetry are accompanied by cell surface exposure of negatively charged phospholipids; we speculate that the Rim101 pathway recognizes the surface charges. INTRODUCTIONBiological membranes are composed of lipid bilayers, and in the plasma membrane the lipid compositions differ between the two leaflets, resulting in asymmetry. Glycerophospholipids and sphingolipids contribute greatly to such asymmetry. Phosphatidylcholine (PC) and complex sphingolipids, including sphingomyelin (SM) and glycosphingolipids in mammals and myo-inositol-containing sphingolipids in the yeast Saccharomyces cerevisiae, are located mainly in the outer (extracytosolic) leaflet. Conversely, phosphatidylserine (PS), phosphatidylethanolamine (PE), and phosphatidylinositol (PI) are confined to the inner (cytosolic) leaflet (Pomorski et al., 2004;Ikeda et al., 2006). The hydrophilic nature of the headgroups of these amphiphilic lipids hinders their ability to traverse the hydrophobic membrane interior, and spontaneous flip-flop of the protein-free model membrane occurs only in low frequency (Bai and Pagano, 1997). However, situated in the biological membranes are enzymes called lipid translocases or flippases, which catalyze the transbilayer movement of lipids and establish their asymmetry. Recent studies have identified three classes of translocases/transporters for glycerophospholipids: ABC transporters, P-type ATPases, and scramblases (Holthuis and Levine, 2005;Ikeda et al., 2006). ABC transporters catalyze flop, which is the movement from the cytosolic to extracytosolic leaflet, whereas P-type ATPases stimulate flip, the reverse movement. Scramblases randomize the lipid distribution between the two leaflets. Sometimes, discriminating between a translocase and transporter is difficult. For example, it is unclear whether a lipid in one leaflet is directly translocated to the other leaflet or is first transported to the other side of the hydrophil...

show abstract

“…22,23,31,[34][35][36][37] We therefore sought to investigate if an exposure of yeast to POA alters the relative levels of PE in the 2 leaflets of the PM. To attain this objective, we used the tetracyclic peptide antibiotic cinnamycin whose specific binding to PE in the extracellular leaflet of the PM is known to cause cell lysis; 36,38 hence, the level of PE in this leaflet of the PM bilayer positively correlates with the sensitivity of yeast to cinnamycin. 22 We found that an exposure of WT cells to POA decreases the sensitivity of yeast to cinnamycin ( Fig.…”

mentioning

confidence: 99%

Mechanism of liponecrosis, a distinct mode of programmed cell death

Richard¹,

Beach²,

Piano³

et al. 2014

Cell Cycle

View full text Add to dashboard Cite

show abstract

A Novel Peptide Probe for Studying the Transbilayer Movement of Phosphatidylethanolamine

Cited by 60 publications

References 0 publications

A Novel Human Phosphatidylethanolamine-binding Protein Resists Tumor Necrosis Factor α-induced Apoptosis by Inhibiting Mitogen-activated Protein Kinase Pathway Activation and Phosphatidylethanolamine Externalization

A Novel Human Phosphatidylethanolamine-binding Protein Resists Tumor Necrosis Factor α-induced Apoptosis by Inhibiting Mitogen-activated Protein Kinase Pathway Activation and Phosphatidylethanolamine Externalization

The Rim101 Pathway Is Involved in Rsb1 Expression Induced by Altered Lipid Asymmetry

Mechanism of liponecrosis, a distinct mode of programmed cell death

Contact Info

Product

Resources

About