Stoicheiometrical proton and potassium ion movements accompanying the absorption of amino acids by the yeast Saccharomyces carlsbergensis

Abstract

1. Proton uptake into the yeast Saccharomyces carlsbergensis, was studied at pH4.5-5.5 in the presence of both antimycin and 2-deoxyglucose to inhibit energy metabolism. Previous work had shown that the cells then absorbed about 20nmol of glycine or l-phenylalanine against a considerable amino acid concentration gradient. The addition of the amino acid immediately stimulated the rate of uptake of protons two- to three-fold. About 2 extra equivalents of H(+) accompanied a given amount of the amino acids into the yeast preparations exposed to the metabolic inhibitors for 2-4min and about 1.2 equivalents after 20min exposure. 2. Analogous observations were made during serial additions of glycine, l-phenylalanine, l-leucine and l-lysine to preparations lacking the metabolic inhibitors and deficient in substrates needed for energy metabolism. In fresh cellular preparations the influx of glycine was then closely coupled to a stimulated flow of 2.1 equiv. of H(+) into the yeast. A similar number of K(+) ions left the cells. About 30% of the extra protons was subsequently ejected from the yeast. Deoxyglucose and antimycin together inhibited the ejection of protons. When the yeast had been fed with glucose energy metabolism was stimulated and almost as many protons as were absorbed with the amino acid were apparently ejected again. 3. Yeast preparations containing Na(+), instead of K(+), as the principal cation absorbed about 1 extra equivalent of H(+) after the addition of phenylalanine, glycine or leucine. This response was not observed in the presence of both deoxyglucose and antimycin. 4. The observations show that H(+) and, in certain circumstances, K(+) are co-substrates in the transport of the amino acids into the yeast. An analogy is drawn with the roles of Na(+) and K(+) as co-substrates in certain mammalian systems. The results lead to various models relating the physical flow of the co-substrate ions on the amino acid carrier to the transduction of chemical energy in an associated ion pump forming part of the mechanism for transporting amino acids into the yeast.