Blockage of ATPase-mediated energy supply inducing metabolic disturbances in algal cells under silver nanoparticles stress

Adenosine triphosphate(ATP)generation of aquatic organisms is often subject to nanoparticles(NPs)stress,involving extensive reprogramming of gene expression and changes in enzyme activity accompanied by metabolic disturbances.However,little is known about the mechanism of energy supply by ATP to regulate the metabolism of aquatic organisms under NPs stress.Here,we selected extensively existing silver nanoparticles(AgNPs)to investigate their implications on ATP generation and relevant metabolic pathways in alga(Chlorella vulgaris).Results showed that ATP content significantly decreased by 94.2%of the control(without AgNPs)in the algal cells at 0.20 mg/L AgNPs,which was mainly attributed to the reduction of chloroplast ATPase activity(81.4%)and the downregulation of ATPase-coding genes atpB and atpH(74.5%-82.8%)in chloroplast.Molecular dynamics simulations demonstrated that AgNPs competed with the binding sites of substrates adenosine diphosphate and inorganic phosphate by forming a stable complex with ATPase subunit beta,potentially resulting in the reduced binding efficiency of substrates.Furthermore,metabolomics analysis proved that the ATP content positively correlated with the content of most differential metabolites such as D-talose,myo-inositol,and L-allothreonine.AgNPs remarkably inhibited ATPinvolving metabolic pathways,including inositol phosphate metabolism,phosphatidylinositol signaling system,glycerophospholipid metabolism,aminoacyl-tRNA biosynthesis,and glutathione metabolism.These results could provide a deep understanding of energy supply in regulating metabolic disturbances under NPs stress.