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.
View Article and Find Full Text PDFNuclear 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.
View Article and Find Full Text PDFThe 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.
View Article and Find Full Text PDFTime-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.
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