Vespa Amino Acid Mixture (VAAM) Enhances Electron and Proton Transport in Mitochondria and Leads to Excessive Reactive Oxygen Production

Abstract

Vespa amino acid mixture (VAAM) is a synthetic blend of amino acids, which has been proven to increase rates of aerobic cellular metabolism by increasing net ATP production in various eukaryotes. The mechanism through which VAAM increases ATP production is unknown; our work suggests that VAAM augments the proton motive force. In order to quantitatively evaluate the effects of VAAM on mitochondrial metabolism, we measured the levels of reactive oxygen species, relative NAD+/NADH levels, caspase activation, mitochondrial membrane integrity, and ATP levels in yeast (Saccharomyces cerevisiae) with differing doses of VAAM at multiple time points. The levels of reactive oxygen species increased within five minutes of the exposure to the lowest dose of VAAM (0.003%, 1% of the recommended dose) and time points beyond five minutes yielded toxic levels and activation of caspase. This was also demonstrated with NAD+/NADH levels, higher levels of NAD+ were observed in the VAAM treated cells, indicating an increase in entry of electrons into the transport chain, leading to the observed increase in ATP production. We attempted to counteract the actions of VAAM by adding α‐tocopherol (an antioxidant) and 2,4‐dinitrophenol (DNP, a proton motive force uncoupler) to the VAAM treatment separately. The antioxidants reduced the production of reactive oxygen species only slightly, and did not significantly lower the production of ATP. Surprisingly, DNP, known to uncouple the proton motive force, was successful in prolonging the life of the cell in the presence of VAAM. This taken in account with the increase in ATP production leads us to believe that VAAM facilitates electron transport leading to enhanced coupling with the activity of ATP synthase.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.