Mass Spectrometric Characterization of Oligomers in Pseudomonas aeruginosa Azurin Solutions

We have employed laser-induced liquid bead ion desorption mass spectroscopy (LILBID MS) to study the solution behavior of Pseudomonas aeruginosa azurin as well as two mutants and corresponding Re-labeled derivatives containing a Re(CO)_(3)(4,7-dimethyl-1,10-phenanthroline)^+ chromophore appended to a surface histidine. LILBID spectra show broad oligomer distributions whose particular patterns depend on the solution composition (pure H_(2)O, 20−30 mM NaCl, 20 and 50 mM NaP_i or NH_(4)P_i at pH = 7). The distribution maximum shifts to smaller oligomers upon decreasing the azurin concentration and increasing the buffer concentration. Oligomerization is less extensive for native azurin than its mutants. The oligomerization propensities of unlabeled and Re-labeled proteins are generally comparable, and only Re126 shows some preference for the dimer that persists even in highly diluted solutions. Peak shifts to higher masses and broadening in 20−50 mM NaP_i confirm strong azurin association with buffer ions and solvation. We have found that LILBID MS reveals the solution behavior of weakly bound nonspecific protein oligomers, clearly distinguishing individual components of the oligomer distribution. Independently, average data on oligomerization and the dependence on solution composition were obtained by time-resolved anisotropy of the Re-label photoluminescence that confirmed relatively long rotation correlation times, 6−30 ns, depending on Re−azurin and solution composition. Labeling proteins with Re-chromophores that have long-lived phosphorescence extends the time scale of anisotropy measurements to hundreds of nanoseconds, thereby opening the way for investigations of large oligomers with long rotation times

Biochemical, kinetic, and spectroscopic characterization of Ruegeria pomeroyi DddW - A mononuclear iron-dependent DMSP lyase

The osmolyte dimethylsulfoniopropionate (DMSP) is a key nutrient in marine environments and its catabolism by bacteria through enzymes known as DMSP lyases generates dimethylsulfide (DMS), a gas of importance in climate regulation, the sulfur cycle, and signaling to higher organisms. Despite the environmental significance of DMSP lyases, little is known about how they function at the mechanistic level. In this study we biochemically characterize DddW, a DMSP lyase from the model roseobacter Ruegeria pomeroyi DSS-3. DddW is a 16.9 kDa enzyme that contains a C-terminal cupin domain and liberates acrylate, a proton, and DMS from the DMSP substrate. Our studies show that as-purified DddW is a metalloenzyme, like the DddQ and DddP DMSP lyases, but contains an iron cofactor. The metal cofactor is essential for DddW DMSP lyase activity since addition of the metal chelator EDTA abolishes its enzymatic activity, as do substitution mutations of key metal-binding residues in the cupin motif (His81, His83, Glu87, and His121). Measurements of metal binding affinity and catalytic activity indicate that Fe(II) is most likely the preferred catalytic metal ion with a nanomolar binding affinity. Stoichiometry studies suggest DddW requires one Fe(II) per monomer. Electronic absorption and electron paramagnetic resonance (EPR) studies show an interaction between NO and Fe(II)-DddW, with NO binding to the EPR silent Fe(II) site giving rise to an EPR active species (g = 4.29, 3.95, 2.00). The change in the rhombicity of the EPR signal is observed in the presence of DMSP, indicating that substrate binds to the iron site without displacing bound NO. This work provides insight into the mechanism of DMSP cleavage catalyzed by DddW

Elemental abundances of Galactic bulge planetary nebulae from optical recombination lines

(abridged) Deep long-slit optical spectrophotometric observations are presented for 25 Galactic bulge planetary nebulae (GBPNe) and 6 Galactic disk planetary nebulae (GDPNe). The spectra, combined with archival ultraviolet spectra obtained with the International Ultraviolet Explorer (IUE) and infrared spectra obtained with the Infrared Space Observatory (ISO), have been used to carry out a detailed plasma diagnostic and element abundance analysis utilizing both collisional excited lines (CELs) and optical recombination lines (ORLs). Comparisons of plasma diagnostic and abundance analysis results obtained from CELs and from ORLs reproduce many of the patterns previously found for GDPNe. In particular we show that the large discrepancies between electron temperatures (Te's) derived from CELs and from ORLs appear to be mainly caused by abnormally low values yielded by recombination lines and/or continua. Similarly, the large discrepancies between heavy element abundances deduced from ORLs and from CELs are largely caused by abnormally high values obtained from ORLs, up to tens of solar in extreme cases. It appears that whatever mechanisms are causing the ubiquitous dichotomy between CELs and ORLs, their main effects are to enhance the emission of ORLs, but hardly affect that of CELs. It seems that heavy element abundances deduced from ORLs may not reflect the bulk composition of the nebula. Rather, our analysis suggests that ORLs of heavy element ions mainly originate from a previously unseen component of plasma of Te's of just a few hundred Kelvin, which is too cool to excite any optical and UV CELs.Comment: 35 pages, 27 figures and 16 tables, accepted for publication in MNRA

Detection of delta Scuti-like pulsation in H254, a pre-main sequence F-type star in IC 348

We present time series observations of intermediate mass PMS stars belonging to the young star cluster IC 348. The new data reveal that a young member of the cluster, H254, undergoes periodic light variations with delta Scuti-like characteristics. This occurrence provides an unambiguous evidence confirming the prediction that intermediate-mass pre-main sequence (PMS) stars should experience this transient instability during their approach to the main-sequence. On the basis of the measured frequency f=7.406 c/d, we are able to constrain the intrinsic stellar parameters of H254 by means of linear, non adiabatic, radial pulsation models. The range of the resulting luminosity and effective temperature permitted by the models is narrower than the observational values. In particular, the pulsation analysis allows to derive an independent estimate of the distance to IC 348 of about 320 pc. Further observations could either confirm the monoperiodic nature of H254 or reveal the presence of other frequencies.Comment: 7 pages, including 7 postscript figures, accepted for publication on A&

The high-mass disk candidates NGC7538IRS1 and NGC7538S

Context: The nature of embedded accretion disks around forming high-mass stars is one of the missing puzzle pieces for a general understanding of the formation of the most massive and luminous stars. Methods: Using the Plateau de Bure Interferometer at 1.36mm wavelengths in its most extended configuration we probe the dust and gas emission at ~0.3",corresponding to linear resolution elements of ~800AU. Results: NGC7538IRS1 remains a single compact and massive gas core with extraordinarily high column densities, corresponding to visual extinctions on the order of 10^5mag, and average densities within the central 2000AU of ~2.1x10^9cm^-3 that have not been measured before. We identify a velocity gradient across in northeast-southwest direction that is consistent with the mid-infrared emission, but we do not find a gradient that corresponds to the proposed CH3OH maser disk. The spectral line data toward NGC7538IRS1 reveal strong blue- and red-shifted absorption toward the mm continuum peak position. The red-shifted absorption allows us to estimate high infall rates on the order of 10^-2 Msun/yr. Although we cannot prove that the gas will be accreted in the end, the data are consistent with ongoing star formation activity in a scaled-up low-mass star formation scenario. Compared to that, NGC7538S fragments in a hierarchical fashion into several sub-sources. While the kinematics of the main mm peak are dominated by the accompanying jet, we find rotational signatures from a secondary peak. Furthermore, strong spectral line differences exist between the sub-sources which is indicative of different evolutionary stages within the same large-scale gas clump.Comment: 15 pages, 12 figures, accepted for A&

Converting NAD83 GPS heights into NAVD88 elevations with LVGEOID, a hybrid geoid height model for the Long Valley volcanic region, California

A GPS survey of leveling benchmarks done in Long Valley Caldera in 1999 showed that the application of the National Geodetic Survey (NGS) geoid model GEOID99 to tie GPS heights to historical leveling measurements would significantly underestimate the caldera ground deformation (known from other geodetic measurements). The NGS geoid model was able to correctly reproduce the shape of the deformation, but required a local adjustment to give a realistic estimate of the magnitude of the uplift. In summer 2006, the U.S. Geological Survey conducted a new leveling survey along two major routes crossing the Long Valley region from north to south (Hwy 395) and from east to west (Hwy 203 – Benton Crossing). At the same time, 25 leveling bench marks were occupied with dual frequency GPS receivers to provide a measurement of the ellipsoid heights. Using the heights from these two surveys, we were able to compute a precise geoid height model (LVGEOID) for the Long Valley volcanic region. Our results show that although the LVGEOID and the latest NGS GEOID03 model practically coincide in areas outside the caldera, there is a difference of up to 0.2 m between the two models within the caldera. Accounting for this difference is critical when using the geoid height model to estimate the ground deformation due to magmatic or tectonic activity in the calder

Some Hermite-Hadamard type inequalities in the class of hyperbolic p-convex functions

In this paper, obtained some new class of Hermite-Hadamard and Hermite-Hadamard-Fejer type inequalities via fractional integrals for the p-hyperbolic convex functions. It is shown that such inequalities are simple consequences of Hermite-Hadamard-Fejer inequality for the p-hyperbolic convex function.Comment: 11 page