26 research outputs found
Ligand Binding Study of Human PEBP1/RKIP: Interaction with Nucleotides and Raf-1 Peptides Evidenced by NMR and Mass Spectrometry
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.METASU
Observations of Ly Emitters at High Redshift
In this series of lectures, I review our observational understanding of
high- Ly emitters (LAEs) and relevant scientific topics. Since the
discovery of LAEs in the late 1990s, more than ten (one) thousand(s) of LAEs
have been identified photometrically (spectroscopically) at to . These large samples of LAEs are useful to address two major astrophysical
issues, galaxy formation and cosmic reionization. Statistical studies have
revealed the general picture of LAEs' physical properties: young stellar
populations, remarkable luminosity function evolutions, compact morphologies,
highly ionized inter-stellar media (ISM) with low metal/dust contents, low
masses of dark-matter halos. Typical LAEs represent low-mass high- galaxies,
high- analogs of dwarf galaxies, some of which are thought to be candidates
of population III galaxies. These observational studies have also pinpointed
rare bright Ly sources extended over kpc, dubbed
Ly blobs, whose physical origins are under debate. LAEs are used as
probes of cosmic reionization history through the Ly damping wing
absorption given by the neutral hydrogen of the inter-galactic medium (IGM),
which complement the cosmic microwave background radiation and 21cm
observations. The low-mass and highly-ionized population of LAEs can be major
sources of cosmic reionization. The budget of ionizing photons for cosmic
reionization has been constrained, although there remain large observational
uncertainties in the parameters. Beyond galaxy formation and cosmic
reionization, several new usages of LAEs for science frontiers have been
suggested such as the distribution of {\sc Hi} gas in the circum-galactic
medium and filaments of large-scale structures. On-going programs and future
telescope projects, such as JWST, ELTs, and SKA, will push the horizons of the
science frontiers.Comment: Lecture notes for `Lyman-alpha as an Astrophysical and Cosmological
Tool', Saas-Fee Advanced Course 46. Verhamme, A., North, P., Cantalupo, S., &
Atek, H. (eds.) --- 147 pages, 103 figures. Abstract abridged. Link to the
lecture program including the video recording and ppt files :
https://obswww.unige.ch/Courses/saas-fee-2016/program.cg
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X-RAY DETECTED ACTIVE GALACTIC NUCLEI IN DWARF GALAXIES AT 0 < z < 1
We present a sample of accreting supermassive black holes (SMBHs) in dwarf galaxies at z < 1. We identify dwarf galaxies in the NEWFIRM Medium Band Survey with stellar masses of M-star < 3 x 10(9) M-circle dot that have spectroscopic redshifts from the DEEP2 survey and lie within the region covered by deep ( flux limit of similar to 5 x 10(-17)-6 x 10(-16) erg cm(-2) s(-1)) archival Chandra X-ray data. From our sample of 605 dwarf galaxies, 10 exhibit X-ray emission consistent with that arising from active galactic nucleus (AGN) activity. If black-hole mass scales roughly with stellar mass, then we expect that these AGNs are powered by SMBHs with masses of similar to 10(5)-10(6) M-circle dot and typical Eddington ratios of similar to 5%. Furthermore, we find an AGN fraction consistent with extrapolations of other searches of similar to 0.6%-3% for 10(9) M-circle dot. M-star <= 3 x 10(9) M-circle dot and 0.1 < z < 0.6. Our AGN fraction is in good agreement with a semi-analytic model, suggesting that, as we search larger volumes, we may use comparisons between observed AGN fractions and models to understand seeding mechanisms in the early universe