Vps3 and Vps8 control integrin trafficking from early to recycling endosomes and regulate integrin-dependent functions

Recycling endosomes maintain plasma membrane homeostasis and are important for cell polarity, migration, and cytokinesis. Yet, the molecular machineries that drive endocytic recycling remain largely unclear. The CORVET complex is a multi-subunit tether required for fusion between early endosomes. Here we show that the CORVET-specific subunits Vps3 and Vps8 also regulate vesicular transport from early to recycling endosomes. Vps3 and Vps8 localise to Rab4-positive recycling vesicles and co-localise with the CHEVI complex on Rab11-positive recycling endosomes. Depletion of Vps3 or Vps8 does not affect transferrin recycling, but delays the delivery of internalised integrins to recycling endosomes and their subsequent return to the plasma membrane. Consequently, Vps3/8 depletion results in defects in integrin-dependent cell adhesion and spreading, focal adhesion formation, and cell migration. These data reveal a role for Vps3 and Vps8 in a specialised recycling pathway important for integrin trafficking

On the electronic structure of electron doped LaOFeAs as seen by X-ray absorption spectroscopy

We investigated the recently found superconductor LaO_{1-x}F_xFeAs by X-ray absorption spectroscopy (XAS). From a comparison of the O K-edge with LDA calculations we find good agreement and are able to explain the structure and changes of the spectra with electron doping. An important result from this edge is a limitation of the Hubbard U to values not significantly larger than 1 eV. From experimental Fe L_2,3-edge spectra and charge transfer multiplet calculations we gain further information on important physical values such as hopping parameters, the charge transfer energy Delta, and the on-site Hubbard U. Furthermore we find the system to be very covalent with a large amount of ligand holes. A shift in the chemical potential is visible in the O K- and Fe L_2,3-edge spectra which emphasizes the importance of band effects in these compounds.Comment: 4 pages, 2 figure

Comparing optimized CO emission estimates using MOPITT or NOAA surface network observations

[1] This paper compares two global inversions to estimate carbon monoxide (CO) emissions for 2004. Either surface flask observations from the National Oceanic and Atmospheric Administration Earth System Research Laboratory (NOAA/ESRL) Global Monitoring Division (GMD) or CO total columns from the Measurement of Pollution in the Troposphere (MOPITT) instrument are assimilated in a 4D-Var framework. Inferred emission estimates from the two inversions are consistent over the Northern Hemisphere (NH). For example, both inversions increase anthropogenic CO emissions over Europe (from 46 to 94 Tg CO/yr) and Asia (from 222 to 420 Tg CO/yr). In the Southern Hemisphere (SH), three important findings are reported. First, due to their different vertical sensitivity, the stations-only inversion increases SH biomass burning emissions by 108 Tg CO/yr more than the MOPITT-only inversion. Conversely, the MOPITT-only inversion results in SH natural emissions (mainly CO from oxidation of NMVOCs) that are 185 Tg CO/yr higher compared to the stations-only inversion. Second, MOPITT-only derived biomass burning emissions are reduced with respect to the prior which is in contrast to previous (inverse) modeling studies. Finally, MOPITT derived total emissions are significantly higher for South America and Africa compared to the stations-only inversion. This is likely due to a positive bias in the MOPITT V4 product. This bias is also apparent from validation with surface stations and ground-truth FTIR columns. Our results show that a combined inversion is promising in the NH. However, implementation of a satellite bias correction scheme is essential to combine both observational data sets in the SH

Clusters in the inner spiral arms of M51: the cluster IMF and the formation history

We study the cluster population in a region of 3.2x3.2 kpc^2 in the inner spiral arms of the intergacting galaxy M51, at a distance of about 1 to 3 kpc from the nucleus, based on HST--WFPC2 images taken through five broadband and two narrowband filters. We found 877 cluster candidates and we derived their ages, initial masses and extinctions by comparing their energy distribution with the Starburst99 cluster models. We describe the 3 and 2-dimensional least-square energy fitting method that was used (3DEF, 2DEF). The lack of [OIII] emission in even the youngest clusters with strong H-alpha emission, indicates the absence of the most massive stars and suggests a mass upper limit of about 25 to 30 solar masses. The mass versus age distribution of the clusters shows a drastic decrease in the number of clusters with age, which indicates that cluster disruption is occurring on a timescale of about 10 Myr for low mass clusters. The cluster initial mass function for clusters younger than 10 Myr has an exponent of alpha = 2.0 (+- 0.05) We derived the cluster formation history from clusters with an initial mass larger than 10^4 solar masses. There is no evidence for a peak in the cluster formation rate within a factor two at about 200 to 400 Myr ago, i.e. at the time of the interaction with the companion galaxy NGC 5194.Comment: 15 pages, 15 figures. Accepted for publication by Astronomy and Astrophysic

Staphylococcus aureus sortase a-mediated incorporation of peptides: Effect of peptide modification on incorporation

The endogenous Staphylococcus aureus sortase A (SrtA) transpeptidase covalently anchors cell wall-anchored (CWA) proteins equipped with a specific recognition motif (LPXTG) into the peptidoglycan layer of the staphylococcal cell wall. Previous in situ experiments have shown that SrtA is also able to incorporate exogenous, fluorescently labelled, synthetic substrates equipped with the LPXTG motif (K(FITC)LPETG-amide) into the bacterial cell wall, albeit at high concentrations of 500 μM to 1 mM. In the present study, we have evaluated the effect of substrate modification on the incorporation efficiency. This revealed that (i) by elongation of LPETG-amide with a sequence of positively charged amino acids, derived from the C-terminal domain of physiological SrtA substrates, the incorporation efficiency was increased by 20-fold at 10 μM, 100 μM and 250 μM; (ii) Substituting aspartic acid (E) for methionine increased the incorporation of the resulting K(FITC)LPMTG-amide approximately three times at all concentrations tested; (iii) conjugation of the lipid II binding antibiotic vancomycin to K(FITC)LPMTG-amide resulted in the same incorporation levels as K(FITC)LPETG-amide, but much more efficient at an impressive 500-fold lower substrate concentration. These newly developed synthetic substrates can potentially find broad applications in for example the in situ imaging of bacteria; the incorporation of antibody recruiting moieties; the targeted delivery and covalent incorporation of antimicrobial compounds into the bacterial cell wall

Inhibition of the Enzymatic Activity of Heme Oxygenases by Azole-Based Antifungal Drugs

ABSTRACT Ketoconazole (KTZ) and other azole antifungal agents are known to have a variety of actions beyond the inhibition of sterol synthesis in fungi. These drugs share structural features with a series of novel heme oxygenase (HO) inhibitors designed in our laboratory. Accordingly, we hypothesized that therapeutically used azole-based antifungal drugs are effective HO inhibitors. Using gas chromatography to quantify carbon monoxide formation in vitro and in vivo, we have shown that azole-containing antifungal drugs are potent HO inhibitors. Terconazole, sulconazole, and KTZ were the most potent drugs with IC 50 values of 0.41 Ϯ 0.01, 1.1 Ϯ 0.4, and 0.3 Ϯ 0.1 M for rat spleen microsomal HO activity, respectively. Kinetic characterization revealed that KTZ was a noncompetitive HO inhibitor. In the presence of KTZ (2.5 and 10 M), K m values for both rat spleen and brain microsomal HO were not altered; however, a significant decrease in the catalytic capacity (V max ) was observed (P Ͻ 0.005). KTZ was also found to weakly inhibit nitric-oxide synthase with an IC 50 of 177 Ϯ 2 M but had no effect on the enzymatic activity of NADPH cytochrome P450 reductase. Because these drugs were effective within the concentration range observed in humans, it is possible that inhibition of HO may play a role in some of the pharmacological actions of these antimycotic drugs

A multi-year methane inversion using SCIAMACHY, accounting for systematic errors using TCCON measurements

This study investigates the use of total column CH₄ (<i>X</i>CH₄) retrievals from the SCIAMACHY satellite instrument for quantifying large-scale emissions of methane. A unique data set from SCIAMACHY is available spanning almost a decade of measurements, covering a period when the global CH₄ growth rate showed a marked transition from stable to increasing mixing ratios. The TM5 4DVAR inverse modelling system has been used to infer CH₄ emissions from a combination of satellite and surface measurements for the period 2003–2010. In contrast to earlier inverse modelling studies, the SCIAMACHY retrievals have been corrected for systematic errors using the TCCON network of ground-based Fourier transform spectrometers. The aim is to further investigate the role of bias correction of satellite data in inversions. Methods for bias correction are discussed, and the sensitivity of the optimized emissions to alternative bias correction functions is quantified. It is found that the use of SCIAMACHY retrievals in TM5 4DVAR increases the estimated inter-annual variability of large-scale fluxes by 22% compared with the use of only surface observations. The difference in global methane emissions between 2-year periods before and after July 2006 is estimated at 27–35 Tg yr⁻¹. The use of SCIAMACHY retrievals causes a shift in the emissions from the extra-tropics to the tropics of 50 ± 25 Tg yr⁻¹. The large uncertainty in this value arises from the uncertainty in the bias correction functions. Using measurements from the HIPPO and BARCA aircraft campaigns, we show that systematic errors in the SCIAMACHY measurements are a main factor limiting the performance of the inversions. To further constrain tropical emissions of methane using current and future satellite missions, extended validation capabilities in the tropics are of critical importance

Anisotropy of 4f states in 3d-4f single-molecule magnets

We have measured angular-dependent fluorescence-yield x-ray magnetic circular dichroism spectra on single crystals of the heterometallic 3d-4f 12-metallacrown-4 TbMn4 and DyMn4 complexes. Simulated spectra using crystal-field multiplet calculations reproduce the experimentally observed spectra. The orientation of the molecules causes linear dichroism spectra of the 4f absorption spectra. This natural linear dichroism shows the anisotropic charge distribution of the rare-earth 4f state in the tetragonal crystal field despite the small 4f crystal-field splitting. The magnetic moment of the molecule is dominated by the rare-earth moment revealing a considerably large contribution of orbital moment. From a sum-rule analysis of experimental and simulated x-ray magnetic circular dichroism, we determined corrected spin and orbital Dy moments at low temperature (14 K) within a magnetic field of 7 T. We find a significant angular dependence of the Dy magnetic moments, indicating the presence of fourth-order magnetic anisotropy