Research Paper Volume 13, Issue 6 pp 7846—7871

Enhancing lifespan of budding yeast by pharmacological lowering of amino acid pools

Myr treatment lowers the rate of amino acid uptake. Uptake of radioactive amino acids was quantified after 4 h of growth of the indicated prototrophic strains (BY4741 background) in SDC medium. (A) The concentration of amino acids in the uptake assay was equal to one-fifth of the concentration in SDC medium. The mixture of 14C-labeled amino acids contained Glu (16%), branched chain amino acids (29%), Ala (13%), Asp (12.5%) and Phe (8.9%) with smaller amounts of the other amino acids. (B) Uptake of 3H-Met was performed with 12 μmol/L substrate, the Km for Mup1 transport, to evaluate activity of this high affinity transporter. (C) Uptake of 3H-Met was performed with 68 μmol/L substrate to determine if the activity of other AATs besides Mup1, with a lower affinity for Met, responded to drug treatment. (D, E). Uptake assays for 14C-glutamate and 14C-arginine were performed in medium containing 100 μmol/L substrate. Statistical significance determined by using the Student’s t-test for all data shown in this Figure and error bars are SD.

Figure 3. Myr treatment lowers the rate of amino acid uptake. Uptake of radioactive amino acids was quantified after 4 h of growth of the indicated prototrophic strains (BY4741 background) in SDC medium. (A) The concentration of amino acids in the uptake assay was equal to one-fifth of the concentration in SDC medium. The mixture of 14C-labeled amino acids contained Glu (16%), branched chain amino acids (29%), Ala (13%), Asp (12.5%) and Phe (8.9%) with smaller amounts of the other amino acids. (B) Uptake of 3H-Met was performed with 12 μmol/L substrate, the Km for Mup1 transport, to evaluate activity of this high affinity transporter. (C) Uptake of 3H-Met was performed with 68 μmol/L substrate to determine if the activity of other AATs besides Mup1, with a lower affinity for Met, responded to drug treatment. (D, E). Uptake assays for 14C-glutamate and 14C-arginine were performed in medium containing 100 μmol/L substrate. Statistical significance determined by using the Student’s t-test for all data shown in this Figure and error bars are SD.