Protein three-dimensional structure dynamically changes in solution depending on the presence of ligands and interacting proteins. Methods for detecting these changes in protein conformation include ‘protein footprinting,’ using mass spectrometry. We describe herein a new technique, PLIMB (Plasma Induced Modification of Biomolecules), that generates µs bursts of hydroxyl radicals from water, to measure changes in protein structure via altered solvent accessibility of amino acid side chains. PLIMB was first benchmarked with model compounds, and then applied to a biological problem, i.e., ligand (EGF) induced changes in the conformation of the external (ecto) domain of Epidermal Growth Factor Receptor (EGFR). Regions in which oxidation decreased upon adding EGF fall along the dimerization interface, consistent with models derived from crystal structures. These results demonstrate that plasma-generated hydroxyl radicals from water can be used to map protein conformational changes, and provide a readily accessible means of studying protein structure in solution.
During plasma processing, low-k dielectrics are exposed to high levels of vacuum ultraviolet (VUV) radiation that can cause severe damage to dielectric materials. The degree and nature of VUV-induced damage depend on the VUV photon energies and fluence. In this work, we examine the VUV-absorption spectrum of low-k organosilicate glass using specular X-ray reflectivity (XRR). Low-k SiCOH films were exposed to synchrotron VUV radiation with energies ranging from 7 to 21 eV, and the density vs. depth profile of the VUV-irradiated films was extracted from fitting the XRR experimental data. The results show that the depth of the VUV-induced damage layer is a function of the photon energy. Between 7 and 11 eV, the depth of the damaged layer decreases sharply from 110 nm to 60 nm and then gradually increases to 85 nm at 21 eV. The maximum VUV absorption in low-k films occurs between 11 and 15 eV. The depth of the damaged layer was found to increase with film porosity.
This work examines the effect of the frequency and peak applied voltage on hydroxyl-radical generation in a dielectric-barrier plasma discharge between a metallic needle electrode and one electrode covered with dielectric. The authors examine a system that can expose up to 96 liquid samples in an automated fashion without human intervention beyond setting the initial software configuration. Then, hydroxyl-radical concentration, measured through coumarin fluorescence, was measured for 5 s plasma exposures generated under different high-voltage conditions with frequencies from 2 to 16 kHz and amplitudes from 4 to 9 kV. Their results show that an increase in frequency and/or applied voltage, within the range prescribed above and the limits of the high-voltage power supply, can yield up to a 150% increase in fluorescence with an equivalent hydroxyl-radical increase. Applications using typical previous methods, such as the Fenton Reaction, are limited in that they continuously generate hydroxyl radicals over millisecond and longer intervals. These results establish the electrical parameters that can now be applied to polymers, like proteins, which show three dimensional structures that are flexible and fluctuate on a microsecond and nanosecond time scale, with hydroxyl-radical generation on this time scale using this device. Additionally, plasma exposures may be optimized for a great variety of proteins, devices and techniques, where hydroxyl-radical generation is of utmost importance, reducing exposure time and potential subjection of samples to harmful side effects.
Porous SiCOH films are of great interest in semiconductor fabrication due to their low-dielectric constant properties. Post-deposition treatments using ultraviolet (UV) light on organosilicate thin films are required to decompose labile pore generators (porogens) and to ensure optimum network formation to improve the electrical and mechanical properties of low-k dielectrics. The goal of this work is to choose the best vacuum-ultraviolet photon energy in conjunction with vacuum ultraviolet (VUV) photons without the need for heating the dielectric to identify those wavelengths that will have the most beneficial effect on improving the dielectric properties and minimizing damage. VUV irradiation between 8.3 and 8.9 eV was found to increase the hardness and elastic modulus of low-k dielectrics at room temperature. Combined with UV exposures of 6.2 eV, it was found that this "UV/VUV curing" process is improved compared with current UV curing. We show that UV/VUV curing can overcome drawbacks of UV curing and improve the properties of dielectrics more efficiently without the need for high-temperature heating of the dielectric.
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