PubMed · 14071375
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M E TUNSTALL. 1963. TRICHLORETHYLENE--ETHER--AIR.. https://doi.org/10.1111/j.1365-2044.1963.tb13571.x
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Solid supported lipid bilayers are rapidly delaminated when drawn through the air/water interface. We have discovered that a close packed monolayer of specifically bound protein prevents this process. The protection mechanism worked in two ways. First, when protein-protected bilayers were drawn through the air/water interface, a thin bulk water layer was visible over the entire bilayer region, thereby preventing air from contacting the surface. Second, a stream of nitrogen was used to remove all bulk water from a protected bilayer, which remained fully intact as determined by fluorescence microscopy. The condition of this dried bilayer was further probed by fluorescence recovery after photobleaching. It was found that lipids were not two-dimensionally mobile in dry air. However, when the bilayer was placed in a humid environment, 91% of the bleached fluorescence signal was recovered, indicating long-range two-dimensional mobility. The diffusion coefficient of lipids under humid conditions was an order of magnitude slower than the same bilayer under water. Protected bilayers could be rehydrated after drying, and their characteristic diffusion coefficient was reestablished. Insights into the mechanism of bilayer preservation were suggested.
The transketolase gene (cbbT) that encodes the Calvin-Benson-Bassham pathway transketolase (CbbT) of Rhodobacter sphaeroides was overexpressed in Escherichia coli and the recombinant protein purified to homogeneity. Like other transketolases, R. sphaeroides CbbT was found to be inactivated in the presence of oxygen. At its optimal pH of 7.8, CbbT displays a specific activity of 37 U/mg, a KR5P of 949 microM, a KXu5P of 11 microM, and a KThDP of 1.8 microM. Cysteine 160, equivalent to Cys159 of the yeast enzyme, is found within the active site and is loosely conserved amongst several sources of transketolase. To investigate the role of cysteine 160 found in the active site of R. sphaeroides CbbT, this residue was targeted for mutagenesis. Cys160 was changed to alanine, serine, aspartate, and glutamate. To compare the effect of these mutations on ThDP binding, spectral techniques were employed in addition to analysis by enzymatic activity. Fluorescence quenching was used to measure both equilibrium binding constants as well as first order rates of binding. The results of these studies indicated that Cys160 played an important and substantial role in cofactor binding, revealing the importance of this loosely conserved residue. In addition, the Cys160 mutants did not appear to alter oxygen-mediated inactivation.
Previous research has indicated that foliar mercury (Hg) flux is bi-directional, with influence from both atmospheric and soil Hg. This work investigated the role of soil and air Hg concentrations on foliar Hg exchange using a single-plant gas-exchange system. The exchange of Hg vapor with aspen seedlings grown in soil Hg concentrations of 0.03+/-0.01, 5.8+/-0.5, and 12.3+/-1.3 microg g(-1) and exposed to atmospheric Hg concentrations of 2.4+/-0.5, 11.0+/-0.9, and 30.4+/-2.2 ng m(-3) was measured. At background atmospheric Hg concentrations of 2.4 ng m(-3), foliage released Hg at all three soil Hg concentrations and fluxes ranged from 1.6 to 5.5 ng/m(2)/h. At higher atmospheric Hg concentrations (>11 ng m(-3)), net deposition to foliage ranged from -9 to -47 ng/m(2)/h, exhibiting increase uptake with higher air Hg concentrations. Fluxes associated with aspen showed an immediate response to changes in atmospheric Hg concentrations. Compensation points, the air concentration where no net flux of Hg vapor occurred, were 3-4 ng m(-3) in the light and 2-3 ng m(-3) in the dark for trees grown in soils of 0.03 and 6 microg g(-1) Hg content, and 5-6 ng m(-3) in the light and 2.5-3.5 ng m(-3) in the dark for trees grown in 12 microg g(-1) Hg soils.