Mechanism and Kinetics of Copper Complexes Binding to the Influenza A M2 S31N and S31N/G34E Channels.

Copper(II) is known to bind in the influenza virus His37 cluster in the homotetrameric M2 proton channel and block the proton current needed for uncoating. Copper complexes based on iminodiacetate also block the M2 proton channel and show reduced cytotoxicity and zebrafish-embryo toxicity. In voltage-clamp oocyte studies using the ubiquitous amantadine-insensitive M2 S31N variant, the current block showed fast and slow phases, in contrast to the single phase found for amantadine block of wild-type M2. Here, we evaluate the mechanism of block by copper adamantyl iminodiacitate and copper cyclooctyl iminodiacitate complexes and address whether the complexes can coordinate with one or more of the His37 imidazoles. The current traces were fitted to parametrized master equations. The energetics of binding and the rate constants suggest that the first step is copper complex binding within the channel, and the slow step in the current block is the formation of a Cu-histidine coordination complex. Solution-phase isothermal titration calorimetry and density functional theory (DFT) calculations indicate that imidazole binds to the copper complexes. Structural optimization using DFT reveals that the complexes fit inside the channel and project the Cu(II) toward the His37 cluster, allowing one imidazole to form a coordination complex with Cu(II). Electrophysiology and DFT studies also show that the complexes block the G34E amantadine-resistant mutant despite some crowding in the binding site by the glutamates.

Transdermal uptake of benzophenone-3 from clothing: comparison of human participant results to model predictions.

BACKGROUND: Models of transdermal uptake of chemicals from clothing have been developed, but not compared with recent human subject experiments. In a well-characterized experiment, participants wore t-shirts pre-dosed with benzophenone-3 (BP-3) and BP-3 and a metabolite were monitored in urine voids. OBJECTIVE: Compare a dynamic model of transdermal uptake from clothing to results of the human subject experiment. METHODS: The model simulating dynamic transdermal uptake from clothing was coupled with direct measurements of the gas phase concentration of benzophenone-3 (BP-3) near the surface of clothing to simulate the conditions of the human subject experiment. RESULTS: The base-case model results were consistent with the those reported for human subjects. The results were moderately sensitive to parameters such as the diffusivity in the stratum corneum (SC), the SC thickness, and SC-air partition coefficient. The model predictions were most sensitive to the clothing fit. Tighter clothing worn during exposure period significantly increased excretion rates but tighter fit "clean" clothing during post-exposure period acts as a sink that reduces transdermal absorption by transferring BP-3 from skin surface lipids to clothing. The shape of the excretion curve was most sensitive to the diffusivity in the SC and clothing fit. SIGNIFICANCE: This research provides further support for clothing as an important mediator of dermal exposure to environmental chemicals.