Local polarity and hydrogen bonding inside the Sec14p phospholipid-binding cavity: high-field multi-frequency electron paramagnetic resonance studies

Sec14p promotes the energy-independent transfer of either phosphatidylinositol (PtdIns) or phosphatidylcholine (PtdCho) between lipid bilayers in vitro and represents the major PtdIns/PtdCho transfer protein in the budding yeast Saccharomyces cerevisiae. Herein, we employ multi-frequency high-field electron paramagnetic resonance (EPR) to analyze the electrostatic and hydrogen-bonding microenvironments for series of doxyl-labeled PtdCho molecules bound by Sec14p in a soluble protein-PtdCho complex. A structurally similar compound, 5-doxyl stearic acid dissolved in a series of solvents, was used for experimental calibration. The experiments yielded two-component rigid limit 130- and 220-GHz EPR spectra with excellent resolution in the gx region. Those components were assigned to hydrogen-bonded and nonhydrogen-bonded nitroxide species. Partially resolved 130-GHz EPR spectra from n-doxyl-PtdCho bound to Sec14p were analyzed using this two-component model and allowed quantification of two parameters. First, the fraction of hydrogen-bonded nitroxide species for each n-doxyl-PtdCho was calculated. Second, the proticity profile along the phospholipid-binding cavity of Sec14p was characterized. The data suggest the polarity gradient inside the Sec14p cavity is a significant contributor to the driving molecular forces for extracting a phospholipid from the bilayer. Finally, the enhanced g-factor resolution of EPR at 130 and 220 GHz provides researchers with a spectroscopic tool to deconvolute two major contributions to the x-component of the nitroxide g-matrix: hydrogen-bond formation and local electrostatic effects.     

Electron paramagnetic resonance study of radiation damage in photosynthetic reaction center crystals

Electron paramagnetic resonance (EPR) was used to simultaneously study radiation-induced cofactor reduction and damaging radical formation in single crystals of the bacterial reaction center (RC). Crystals of Fe-removed/Zn-replaced RC protein from Rhodobacter ( R.) sphaeroides R26 were irradiated with varied radiation doses at cryogenic temperature and analyzed for radiation-induced free radical formation and alteration of light-induced photosynthetic electron transfer activity using high-field (HF) D-band (130 GHz) and X-band (9.5 GHz) EPR spectroscopies. These analyses show that the formation of radiation-induced free radicals saturated at doses 1 order of magnitude smaller than the amount of radiation at which protein crystals lose their diffraction quality, while light-induced RC activity was found to be lost at radiation doses at least 1 order of magnitude lower than the dose at which radiation-induced radicals exhibited saturation. HF D-band EPR spectra provide direct evidence for radiation-induced reduction of the quinones and possibly other cofactors. These results demonstrate that substantial radiation damage is likely to have occurred during X-ray diffraction data collection used for photosynthetic RC structure determination. Thus, both radiation-induced loss of photochemical activity in RC crystals and reduction of the quinones are important factors that must be considered when correlating spectroscopic and crystallographic measurements of quinone site structures.     

Evening chronotype,late weekend sleep times and social jetlag as possible causes of sleep curtailment after maintaining perennial DST: ain’t they as black as they are painted?

ABSTRACT

People sleep less in response to setting social clocks earlier relative to the sun clocks. We proposed here a model-based approach for estimating sleep loss as the difference between weekend and weekday risetimes divided on the difference between weekend risetime and weekday bedtime. We compared this approach with a traditional approach to estimating sleep curtailment as the difference in weekly average sleep duration in two conditions. Weekday and weekend sleep times reported for 320 samples provided possibility of testing whether evening types with later weekend sleep times and larger social jetlag differ from morning types with earlier weekend sleep times and smaller social jetlag on amount of sleep lost (1) throughout the week and (2) in response to an advance of weekday wakeups, for instance, after the expected installation of perennial Daylight Saving Time (DST). We found that (1) an amount of sleep lost due to advancing shift of weekday wakeups depends upon neither chronotype nor weekend sleep times nor social jetlag, (2) a very large amount of sleep is usually lost by evening types with later weekend sleep times and larger social jetlag and (3) an essential sleep loss is caused by our usual work/school schedules, even in morning types with early weekend sleep times and small social jetlag. As compared to such permanent sleep losses experienced by any types, an additional loss due to switching from Standard Time (ST) to perennial DST are expected to be relatively small. We also found that the traditional way of calculation of sleep curtailment leads to paradoxical conclusions, such as (1) sleep loss is larger when social jetlag is smaller, not larger, (2) sleep loss is larger when weekend sleep times are earlier, not later, (3) despite 1-h difference between two student samples in weekday wakeups, their sleep losses can be identical.     

The Evolution of Human Probiotics: Challenges and Prospects

Probiotics and Antimicrobial Proteins - In recent years, the intestinal microbiota has been found to greatly influence a number of biological processes important for human health and longevity....     

Ionization of sputtered metal atoms in a microwave ECR plasma source

The ionization of sputtered aluminum atoms in the plasma of a microwave ECR discharge intended for metal coating of submicron-size structures in microelectronics is studied. The spatial distributions of xenon plasma parameters and their variations under the action of metal atoms are investigated using probe and optical emission spectroscopy techniques.     

Ionization fraction of the sputtered metal flux in a hollow cathode magnetron

Results from studies of the parameters of a novel type of plasma source—a hollow cathode magnetron—are presented. The magnetron operates at a gas pressure of 5–20 mTorr, the discharge power being in the range of 0.5–4 kW. At discharge powers exceeding 2 kW, a plasma flow with a density of higher than 10¹¹ cm^?3 and length of up to 30 cm forms at the magnetron output. Using a grid quartz crystal microbalance, the ionized copper flux fraction was measured as a function of the gas pressure, discharge power, and distance from the target. At gas pressures of higher than 15 mTorr, the degree of ionization at a distance of 31 cm exceeds 50%.