Cystic fibrosis transmembrane conductance regulator (CFTR): closed and open state channel models.

The cystic fibrosis transmembrane conductance regulator (CFTR) is a member of the ATP binding cassette (ABC) transporter superfamily. CFTR controls the flow of anions through the apical membrane of epithelia. Dysfunctional CFTR causes the common lethal genetic disease cystic fibrosis. Transitions between open and closed states of CFTR are regulated by ATP binding and hydrolysis on the cystosolic nucleotide binding domains (NBDs), which are coupled with the transmembrane domains (TMDs) forming the pathway for anion permeation. Lack of structural data hampers a global understanding of CFTR, and thus the development of rational approaches directly targeting defective CFTR. In this work, we explored possible conformational states of the CFTR gating cycle by means of homology modeling. As templates, we used structures of homologous ABC transporters, namely TM287- 288, ABC-B10, McjD and Sav1866. In the light of published experimental results, structural analysis of the transmembrane cavity suggests that the TM287-288-based CFTR model could correspond to a commonly occupied closed state, while the McjD-based model could represent an open state. The models capture the important role played by Phe337 as a filter/gating residue and provide structural information on the conformational transition from closed to open channel

Mutant cycles at CFTR's non-canonical ATP-binding site support little interface separation during gating

Cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride channel belonging to the adenosine triphosphate (ATP)-binding cassette (ABC) superfamily. ABC proteins share a common molecular mechanism that couples ATP binding and hydrolysis at two nucleotide-binding domains (NBDs) to diverse functions. This involves formation of NBD dimers, with ATP bound at two composite interfacial sites. In CFTR, intramolecular NBD dimerization is coupled to channel opening. Channel closing is triggered by hydrolysis of the ATP molecule bound at composite site 2. Site 1, which is non-canonical, binds nucleotide tightly but is not hydrolytic. Recently, based on kinetic arguments, it was suggested that this site remains closed for several gating cycles. To investigate movements at site 1 by an independent technique, we studied changes in thermodynamic coupling between pairs of residues on opposite sides of this site. The chosen targets are likely to interact based on both phylogenetic analysis and closeness on structural models. First, we mutated T460 in NBD1 and L1353 in NBD2 (the corresponding site-2 residues become energetically coupled as channels open). Mutation T460S accelerated closure in hydrolytic conditions and in the nonhydrolytic K1250R background; mutation L1353M did not affect these rates. Analysis of the double mutant showed additive effects of mutations, suggesting that energetic coupling between the two residues remains unchanged during the gating cycle. We next investigated pairs 460-1348 and 460-1375. Although both mutations H1348A and H1375A produced dramatic changes in hydrolytic and nonhydrolytic channel closing rates, in the corresponding double mutants these changes proved mostly additive with those caused by mutation T460S, suggesting little change in energetic coupling between either positions 460-1348 or positions 460-1375 during gating. These results provide independent support for a gating model in which ATP-bound composite site 1 remains closed throughout the gating cycle

Structure of Transmembrane Helix 8 and Possible Membrane Defects in CFTR

The cystic fibrosis transmembrane conductance regulator (CFTR) is an ion channel that regulates the flow of anions across epithelia. Mutations in CFTR cause cystic fibrosis. CFTR belongs to the ATP-binding cassette transporter superfamily, and gating is controlled by phosphorylation and ATP binding and hydrolysis. Recently obtained ATP-free and ATP-bound structures of zebrafish CFTR revealed an unwound segment of transmembrane helix (TM) 8, which appears to be a unique feature of CFTR not present in other ATP-binding cassette transporter structures. Here, using μs-long molecular dynamics simulations, we investigate the interactions formed by this TM8 segment with nearby helices in both ATP-free and ATP-bound states. We highlight ATP-dependent interactions as well as the structural role of TM8 in maintaining the functional architecture of the pore via interactions common to both the ATP-bound and ATP-free state. The results of the molecular dynamics simulations are discussed in the context of the gating mechanism of CFTR

Tailored Torsion and Bending-Resistant Avian-Inspired Structures

The escalating demand for torsion- and bending-resistant structures paired with the need for more efficient use of materials and geometries, have led to novel bio-inspired ingenious solutions. However, lessons from Nature could be as inspiring as they are puzzling: plants and animals offer an enormous range of promising but hierarchically complex configurations. Avian bones are prominent candidates for addressing the torsional and bending issue. They present a unique intertwining of simple components: helicoidal ridges and crisscrossing struts, able to bear flexural and twisting actions of winds. Here, it is set how to harmonically move from the natural to the engineering level to formalize and analyze the biological phenomena under controlled design conditions. The effect of ridges and struts is isolated and combined toward tailored torsion and bending-resistant arrangements. Then the biological level is revisited to extrapolate the avian allometric design approach and is translated into multiscale lightweight structures at the engineering level. This study exploits the complexity of Nature and the scalability that characterizes the evolutionary design of bird bones through the design and fabrication versatility allowed by additive manufacturing technologies. This paves the way for exploring the transferability of the proposed solution at multiple engineering scales

Integral field spectroscopy with SINFONI of VVDS galaxies. II. The mass-metallicity relation at 1.2 < z < 1.6

This work aims to provide a first insight into the mass-metallicity (MZ) relation of star-forming galaxies at redshift z~1.4. To reach this goal, we present a first set of nine VVDS galaxies observed with the NIR integral-field spectrograph SINFONI on the VLT. Oxygen abundances are derived from empirical indicators based on the ratio between strong nebular emission-lines (Halpha, [NII]6584 and [SII]6717,6731). Stellar masses are deduced from SED fitting with Charlot & Bruzual (2007) population synthesis models, and star formation rates are derived from [OII]3727 and Halpha emission-line luminosities. We find a typical shift of 0.2-0.4 dex towards lower metallicities for the z~1.4 galaxies, compared to the MZ-relation in the local universe as derived from SDSS data. However, this small sample of eight galaxies does not show any clear correlation between stellar mass and metallicity, unlike other larger samples at different redshift (z~0, z~0.7, and z~2). Indeed, our galaxies lie just under the relation at z~2 and show a small trend for more massive galaxies to be more metallic (~0.1 logarithmic slope). There are two possible explanations to account for these observations. First, the most massive galaxies present higher specific star formation rates when compared to the global VVDS sample which could explain the particularly low metallicity of these galaxies as already shown in the SDSS sample. Second, inflow of metal-poor gas due to tidal interactions could also explain the low metallicity of these galaxies as two of these three galaxies show clear signatures of merging in their velocity fields. Finally, we find that the metallicity of 4 galaxies is lower by ~0.2 to 0.4 dex if we take into account the N/O abundance ratio in their metallicity estimate.Comment: 7 pages, 4 figures, accepted in A&A Comments: Comments: more accurate results with better stellar mass estimate

Effective absorbing column density in the gamma-ray burst afterglow X-ray spectra

We investigate the scaling relation between the observed amount of absorption in the X-ray spectra of Gamma Ray Burst (GRB) afterglows and the absorber redshift. Through dedicated numerical simulations of an ideal instrument, we establish that this dependence has a power law shape with index 2.4. However, for real instruments, this value depends on their low energy cut-off, spectral resolution and on the detector spectral response in general. We thus provide appropriate scaling laws for specific instruments. Finally, we discuss the possibility to measure the absorber redshift from X-ray data alone. We find that 10^5-10^6 counts in the 0.3-10 keV band are needed to constrain the redshift with 10% accuracy. As a test case we discuss the XMM-Newton observation of GRB 090618 at z=0.54. We are able to recover the correct redshift of this burst with the expected accuracy.Comment: MNRAS accepted. 6 figures. 3 table

Fatigue Behavior Of Pipeline Steel Under Hydrogen Environment And Low Temperature

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