The molecular basis of Abelson kinase regulation by its αI-helix.

Abelson tyrosine kinase (Abl) is regulated by the arrangement of its regulatory core, consisting sequentially of the SH3, SH2, and kinase (KD) domains, where an assembled or disassembled core corresponds to low or high kinase activity, respectively. It was recently established that binding of type II ATP site inhibitors, such as imatinib, generates a force from the KD N-lobe onto the SH3 domain and in consequence disassembles the core. Here, we demonstrate that the C-terminal αI-helix exerts an additional force toward the SH2 domain, which correlates both with kinase activity and type II inhibitor-induced disassembly.

The key role of glutamine for protein expression and isotopic labeling in insect cells.

Nuclear magnetic resonance studies of many physiologically important proteins have long been impeded by the necessity to express such proteins in isotope-labeled form in higher eukaryotic cells and the concomitant high costs of providing isotope-labeled amino acids in the growth medium. Economical routes use isotope-labeled yeast or algae extracts but still require expensive isotope-labeled glutamine. Here, we have systematically quantified the effect of N-glutamine on the expression and isotope labeling of different proteins in insect cells.

Time-resolved excited state energetics of the solvated electron in sodium-doped water clusters.

The energetics and dynamics of the first electronically excited state of solvated electron in sodium-doped water clusters has been studied, by means of time-resolved electron spectra created in a pump-probe fs-laser experiment. The Na ··· (H2O)n clusters were excited by pulses at a wavelength of 795 nm, while ionization was achieved at a wavelength of 398 nm, and the overall cross-correlation fwhm was about 50 fs. Mass-resolved electron spectra were taken using photoelectron-photoion coincidence (PEPICO) spectroscopy for cluster sizes ranging from n = 1 up to 22.

Role of alkali cations for the excited state dynamics of liquid water near the surface.

Time-resolved liquid jet photoelectron spectroscopy was used to explore the excited state dynamics at the liquid water surface in the presence of alkali cations. The data were evaluated with the help of ab initio calculations on alkali-water clusters and an extension of these results on the basis of the dielectric continuum model: 160 nm, sub-20 fs vacuum ultraviolet pulses excite water molecules in the solvent shell of Na(+) or K(+) cations and evolve into a transient hydrated complex of alkali-ion and electron. The vertical ionization energy of this transient is about 2.

Time-resolved imaging of laser-induced refractive index changes in transparent media.

We describe a method to visualize ultrafast laser-induced refractive index changes in transparent materials with a 310 fs impulse response and a submicrometer spatial resolution. The temporal profile of the laser excitation sequence can be arbitrarily set on the subpicosecond and picosecond time scales with a pulse shaping unit, allowing for complex laser excitation. Time-resolved phase contrast microscopy reveals the real part of the refractive index change and complementary time-resolved optical transmission microscopy measurements give access to the imaginary part of the refractive index in the irradiated region.

Changing nucleotide specificity of the DEAD-box helicase Hera abrogates communication between the Q-motif and the P-loop.

DEAD-box proteins disrupt or remodel RNA and protein/RNA complexes at the expense of ATP. The catalytic core is composed of two flexibly connected RecA-like domains. The N-terminal domain contains most of the motifs involved in nucleotide binding and serves as a minimalistic model for helicase/nucleotide interactions.

Ultrafast photo-excitation dynamics in isolated, neutral water clusters.

Using the efficient nonlinear conversion scheme which was recently developed in our group [M. Beutler, M. Ghotbi, F.

Single-pulse ultrafast laser imprinting of axial dot arrays in bulk glasses.

Ultrafast laser processing of bulk transparent materials can significantly gain flexibility when the number of machining spots is increased. We present a photoinscription regime in which an array of regular dots is generated before the region of main laser focus under single-pulse exposure in fused silica and borosilicate crown glass without any external spatial phase modulation. The specific position of the dots does not rely on nonlinear propagation effects but is mainly determined by beam truncation and is explained by a Fresnel propagation formalism taking into account beam apodization and linear wavefront distortions at the air/glass interface.

Time-resolved photoelectron spectroscopy of solvated electrons in aqueous NaI solution.

Time-resolved photoelectron spectroscopy was used to study the energetics and dynamics of solvated electrons in aqueous solution. Solvated electrons are generated by ultrafast photodetachment in a 100 mM aqueous NaI solution. Initially, an ensemble of strongly bound ("cold") solvated electrons and an ensemble of weakly bound ("hot") electrons in an unequilibrated solvent environment are observed.

Generation of sub-50-fs vacuum ultraviolet pulses by four-wave mixing in argon.

We report on the generation of femtosecond pulses at 160 nm with energies up to 240 nJ at 1 kHz repetition rate and sub-50-fs pulse duration. This pulse energy is a 1-order-of-magnitude improvement compared with previous sub-100-fs sources in this wavelength range. The pulses are generated by four-wave difference-frequency mixing process between the fundamental of a Ti:sapphire laser and its third harmonic in argon.

Ultrafast dynamics in Na-doped water clusters and the solvated electron.

The lifetimes of the first electronically excited state of (H(2)O)(n)...

Excited-state dynamics of cytosine tautomers.

We report the relaxation dynamics of keto and enol or keto-imino cytosine, photoexcited in the wavelength range of 260-290 nm. Three transients with femtosecond to hundreds of picoseconds lifetimes are observed for the biologically relevant keto tautomer and are assigned to internal conversion and excited-state tautomerization. Only two transients with femtosecond and picosecond lifetimes are identified for the enol or keto-imino tautomer and are assigned to internal conversion processes.

Dynamic ultrafast laser spatial tailoring for parallel micromachining of photonic devices in transparent materials.

Femtosecond laser processing of bulk transparent materials can generate localized positive changes of the refractive index. Thus, by translation of the laser spot, light-guiding structures are achievable in three dimensions. Increasing the number of laser processing spots can consequently reduce the machining effort.

Fragmentation and ionization dynamics of C60 in elliptically polarized femtosecond laser fields.

Ionization and fragmentation of C60 fullerenes is studied in elliptically polarized, intense fs laser fields at 797 nm [I=(0.5-4.3)x10;{14} W cm;{-2}] and contrasted with Xe+, utilizing time-of-flight mass spectrometry.

Ultrafast energy redistribution in C(60) fullerenes: a real time study by two-color femtosecond spectroscopy.

Strong-field excitation and energy redistribution dynamics of C(60) fullerenes are studied by means of time-resolved mass spectrometry in a two-color femtosecond pump-probe setup. Resonant pre-excitation of the electronic system via the first dipole-allowed HOMO-->LUMO+1(t(1g)) (HOMO denotes highest occupied molecular orbital and LUMO denotes lowest unoccupied molecular orbital) transition with ultrashort 25 fs pulses at 399 nm of some 10(12) W cm(-2) results in a highly nonequilibrium distribution of excited electrons and vibrational modes in the neutral species. The subsequent coupling among the electronic and nuclear degrees of freedom is monitored by probing the system with time-delayed 27 fs pulses at 797 nm of some 10(13) W cm(-2).

Interaction between liquid water and hydroxide revealed by core-hole de-excitation.

The hydroxide ion plays an important role in many chemical and biochemical processes in aqueous solution. But our molecular-level understanding of its unusual and fast transport in water, and of the solvation patterns that allow fast transport, is far from complete. One proposal seeks to explain the properties and behaviour of the hydroxide ion by essentially regarding it as a water molecule that is missing a proton, and by inferring transport mechanisms and hydration structures from those of the excess proton.

Pseudoequivalent nitrogen atoms in aqueous imidazole distinguished by chemical shifts in photoelectron spectroscopy.

The photoelectron spectra of aqueous imidazole are presented, and the N 1s and C 1s binding energies are assigned aided by density functional theory calculations. The chemical equivalency of the two nitrogens of the cationic form is directly identified by the occurrence of a single N 1s photoelectron peak, which results from the delocalization of the positive charge over the molecule as a consequence of the Cv symmetry of the system. In contrast to NMR measurements, the photoemission process is faster than the rapid proton exchange in the aqueous environment, making the pseudoequivalent nitrogens of the neutral state clearly distinguishable with a N 1s binding energy shift of 1.

Electron dynamics in charge-transfer-to-solvent states of aqueous chloride revealed by Cl- 2p resonant Auger-electron spectroscopy.

Charge-transfer-to-solvent (CTTS) excited states of aqueous chloride are studied by a novel experimental approach based on resonant inner-shell photoexcitation, Cl(-)aq 2p --> e(i), i = 1-4, which denotes a series of excitations to lowest and higher CTTS states. These states are clearly identified through the occurrence of characteristic spectator Auger decays to double Cl 3p valence-hole states, where the CTTS states can be more stabilized as compared to single Cl(-)aq 2p core excitations and optical valence excitations. Furthermore, we have found for the first time that the CTTS electron e(i) bound by a single Cl 2p hole not only behaves as a spectator e(i) --> e'(i), bound by a double valence-hole state before relaxation of the excited electron (i) itself, but also shows electron dynamics to the relaxed lowest state, e(i) --> e'(1*).

Coherent control of bond breaking in amino acid complexes with tailored femtosecond pulses.

Intense femtosecond laser pulses, judiciously tailored in an adaptive, optimal control feedback loop were used to break preferentially the acyl-N ("peptide") bond of Ac-Phe-NHMe that may be regarded as a dipeptide model. We show that coherent excitation of complex wave packets in the strong-field regime allows to cleave strong backbone bonds in the molecular system preferentially, while keeping other more labile bonds intact. These results show the potential of pulse shaping as a powerful complementary analytical tool for protein sequencing of large biopolymers in addition to the well-known mass spectrometry and chemical analysis.

Electronic structure of adenine and thymine base pairs studied by femtosecond electron-ion coincidence spectroscopy.

Femtosecond pump-probe spectroscopy was combined with photoelectron-photoion coincidence detection to investigate the electronic structure and dynamics of isolated adenine (A) and thymine (T) dimers and the adenine-thymine (AT) base pair. The photoelectron spectra show that pipi* and npi* states are only weakly perturbed in the hydrogen-bound dimers as compared to the monomers. For cationic base pairs with internal energies greater than 1 eV, we observed considerable cluster fragmentation into protonated monomers.

Segmentation of neck lymph nodes in CT datasets with stable 3D mass-spring models segmentation of neck lymph nodes.

Rationale And Objectives: The quantitative assessment of neck lymph nodes in the context of malignant tumors requires an efficient segmentation technique for lymph nodes in tomographic three-dimensional (3D) datasets. We present a stable 3D mass-spring model for lymph node segmentation in computed tomography (CT) datasets.

Materials And Methods: For the first time our model concurrently represents the characteristic gray value range, directed contour information, and shape knowledge, which leads to a robust and efficient segmentation process.

pH-induced protonation of lysine in aqueous solution causes chemical shifts in X-ray photoelectron spectroscopy.

We demonstrate the applicability of X-ray photoelectron spectroscopy to obtain charge- and site-specific electronic structural information of biomolecules in aqueous solution. Changing the pH of an aqueous solution of lysine from basic to acidic results in nitrogen 1s and carbon 1s chemical shifts to higher binding energies. These shifts are associated with the sequential protonation of the two amino groups, which affects both charge state and hydrogen bonding to the surrounding water molecules.

Hydrogen bonding in liquid water probed by resonant Auger-electron spectroscopy.

We have measured resonant and off-resonant Auger-electron spectra of liquid water. Continuumlike transitions near and above the O1s vertical ionization energy are identified by the characteristic normal Auger-electron spectra. On the contrary, well-resolved spectator shifts of the main Auger-electron peak are observed at the liquid-water O1s absorption main edge and near the absorption pre-edge.

Hydrogen bonds in liquid water studied by photoelectron spectroscopy.

The authors report on photoelectron emission spectroscopy measurements of the oxygen 1s orbital of liquid water, using a liquid microjet in ultrahigh vacuum. By suitably changing the soft x-ray photon energy, within 600-1200 eV, the electron probing depth can be considerably altered as to either predominantly access the surface or predominantly bulk water molecules. The absolute probing depth in liquid water was inferred from the evolution of the O1s signal and from comparison with aqueous salt solution.