The small molecule IACS-010759 has been reported to potently inhibit the proliferation of glycolysis-deficient hypoxic tumor cells by interfering with the functions of mitochondrial NADH-ubiquinone oxidoreductase (complex I) without exhibiting cytotoxicity at tolerated doses in normal cells. Considering the significant cytotoxicity of conventional quinone-site inhibitors of complex I, such as piericidin and acetogenin families, we hypothesized that the mechanism of action of IACS-010759 on complex I differs from that of other known quinone-site inhibitors. To test this possibility, here we investigated IACS-010759's mechanism in bovine heart submitochondrial particles. We found that IACS-010759, like known quinone-site inhibitors, suppresses chemical modification by the tosyl reagent AL1 of Asp160 in the 49-kDa subunit, located deep in the interior of a previously proposed quinone-access channel. However, contrary to the other inhibitors, IACS-010759 direction-dependently inhibited forward and reverse electron transfer and did not suppress binding of the quinazoline-type inhibitor [125I]AzQ to the N terminus of the 49-kDa subunit. Photoaffinity labeling experiments revealed that the photoreactive derivative [125I]IACS-010759-PD1 binds to the middle of the membrane subunit ND1 and that inhibitors that bind to the 49-kDa or PSST subunit cannot suppress the binding. We conclude that IACS-010759's binding location in complex I differs from that of any other known inhibitor of the enzyme. Our findings, along with those from previous study, reveal that the mechanisms of action of complex I inhibitors with widely different chemical properties are more diverse than can be accounted for by the quinone-access channel model proposed by structural biology studies.

The effects of vertical confinement on a turbulent shear layer are investigated with large-eddy simulations of a freely developing shear layer (FSL) and a wall-confined shear layer (WSL) that develops between two horizontal walls. In the case of the WSL, the growth of the shear layer is inhibited by the walls. Once the walls prevent the development of the shear layer, highly anisotropic velocity fluctuations become prominent in the flow. These anisotropic velocity fluctuations are recognized as elongated large-scale structures (ELSS), whose streamwise length is much larger than the length scales in the other directions. Spectral analysis confirms that the turbulent kinetic energy is dominated by the ELSS, whose streamwise length grows continuously. A proper orthogonal decomposition can effectively extract a velocity component associated with the ELSS. The isotropy of the Reynolds stress tensor is changed by the presence of the ELSS. These changes in flow characteristics due to the ELSS are not observed in the FSL, where the shear layer thickness increases continuously. These behaviors of the WSL are consistent with those of stably stratified shear layers (SSSLs), where flow structures similar to ELSS also develop when the vertical flow development is confined by the stable stratification. The vertical confinement by the walls or stable stratification strengthens mean shear effects. The flow behavior at large scales in the WSL and SSSL is consistent with rapid distortion theory for turbulence subject to mean shear, suggesting that the development of ELSS is caused by the mean shear.

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