L-Proline transport in Saccharomyces cerevisiae.
Transport of L-proline into Saccharomyces cerevisiae K is mediated by two systems, one with a KT of 31 microM and Jmax of 40 nmol . s-1 . (g dry wt.)-1, the other with KT greater than 2.5 mM and Jmax of 150-165 nmol . s-1 . (g dry wt.)-1. The kinetic properties of the high-affinity system were studied in detail. It proved to be highly specific, the only potent competitive inhibitors being (i) L-proline and its analogs L-azetidine-2-carboxylic acid, sarcosine, D-proline and 3,4-dehydro-DL-proline, and (ii) L-alanine. The other amino acids tested behaved as noncompetitive inhibitors. The high-affinity system is active, has a sharp pH optimum at 5.8-5.9 and, in an Arrhenius plot, exhibits two inflection points at 15 degrees C and 20-21 degrees C. It is trans-inhibited by most amino acids (but probably only the natural substrates act in a trans-noncompetitive manner) and its activity depends to a considerable extent on growth conditions. In cells grown in a rich medium with yeast extract maximum activity is attained during the stationary phase, on a poor medium it is maximal during the early exponential phase. Some 50-60% of accumulated L-proline can leave cells in 90 min (and more if washing is done repeatedly), the efflux being insensitive to 0.5 mM 2,4-dinitrophenol and uranyl ions, the pH between 3 and 7.3, as well as to the presence of 10-100 mM unlabeled L-proline in the outside medium. Its rate and extent are increased by 1% D-glucose and by 10 micrograms nystatin per ml.
D019297:2,4-Dinitrophenol; D000409:Alanine / Q000378:metabolism; D001692:Biological Transport; D004140:Dinitrophenols / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D011392:Proline / Q000031:analogs & derivatives / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000254:growth & development / Q000378:metabolism*; D013696:Temperature
D004140:Dinitrophenols; D011392:Proline; D000409:Alanine; D019297:2,4-Dinitrophenol