Soni Shaikh*, Suman K Nandy, Carles Canti and Sergio Lavandero Pages 1 - 10 ( 10 )
Objective:Bafilomycin-A1 and ML9 are lysosomotropic agents, irrespective of cell types. However, the mechanisms of lysosome targeting of either bafilomycin-A1 or ML9 are unclear.Methods:The present research has been carried out by different molecular and biochemical analyses like western blot, confocal imaging and FACS studies, as well as molecular docking. Results:Our data shows that pre-incubation of neonatal cardiomyocytes with ML9 for 4 h induced cell death, whereas a longer period of time (24h) with bafilomycin-A1 was required to induce an equivalent effect. Neither changes in ROS nor ATP production were associated with such death mechanisms. Flow cytometry, LC3-II expression levels, and LC3-GFP puncta formation revealed a similar lysosomotropic effect for both compounds. We used a molecular docking approach, that predicts a stronger inhibitory activity against V-ATPase-C1 and C2 domains for bafilomycin-A1 in comparison to ML9. Conclusion: Bafilomycin-A1 and ML9 are lysosomotropic agents, involved in cell death events. But such death events are not associated with ATP and ROS production. Furthermore, both drugs target lysosomes by different mechanisms. For the latter, cell death is likely due to lysosomal membrane permeabilization and release of lysosomal proteases into the cytosol.
ML9, Bafilomycin-A1, Lysosome, Cardiomyocyte, Na+/K+-ATPase, Protein modelling, macroautophagy.
Institut de Recerca de Biomedica de Lleida Fundacio Dr. Pifarre (IRBLleida), Lleida, Bioinformatics Infrastructure Facility (BIF), North-Eastern Hill University (NEHU), Tura Campus, Tura, Meghalaya, Institut de Recerca de Biomedica de Lleida Fundacio Dr. Pifarre (IRBLleida), Lleida, Advanced Center for Chronic Disease (ACCDiS), Facultad de Ciencias Químicas y Farmacéuticas & Facultad de Medicina, Universidad de Chile, Santiago