Sodium-dependent transport of branched-chain amino acids by a monensin-sensitive ruminal peptostreptococcus.
A recently isolated ruminal peptostreptococcus which produced large amounts of branched-chain volatile fatty acids grew rapidly with leucine as an energy source in the presence but not the absence of Na. Leucine transport could be driven by an artificial membrane potential (delta psi) only when Na was available, and a chemical gradient of Na+ (delta uNa+) also drove uptake. Because Na+ was taken up with leucine and a Z delta pH could not serve as a driving force (with or without Na), it appeared that leucine was transported in symport with Na+. The leucine carrier could use Li as well as Na and had a single binding site for Na+. The Km for Na was 5.2 mM, and the Km and Vmax for leucine were 77 microM and 328 nmol/mg of protein per min, respectively. Since valine and isoleucine competitively inhibited (Kis of 90 and 49 microM, respectively) leucine transport, it appeared that the peptostreptococcus used a common carrier for branched-chain amino acids. Valine or isoleucine was taken up rapidly, but little ammonia was produced if they were provided individually. The lack of ammonia could be explained by an accumulation of reducing equivalents. The ionophore, monensin, inhibited growth, but leucine was taken up and deaminated at a slow rate. Monensin caused a loss of K, an increase in Na, a slight increase in delta psi, and a decrease in intracellular pH. The inhibition of growth was consistent with a large decrease in ATP.(ABSTRACT TRUNCATED AT 250 WORDS)
Chen
G J
GJ
Department of Animal Science, Cornell University, Ithaca, New York.
Russell
J B
JB
D000597:Amino Acids, Branched-Chain / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D001692:Biological Transport; D002417:Cattle; D006863:Hydrogen-Ion Concentration; D007532:Isoleucine / Q000378:metabolism; D007700:Kinetics; D007930:Leucine / Q000378:metabolism; D008564:Membrane Potentials; D008985:Monensin / Q000494:pharmacology*; D010462:Peptostreptococcus / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism*; D011188:Potassium / Q000378:metabolism; D012417:Rumen / Q000382:microbiology*; D012964:Sodium / Q000494:pharmacology*; D014633:Valine / Q000378:metabolism
D000597:Amino Acids, Branched-Chain; D007532:Isoleucine; D008985:Monensin; D012964:Sodium; D007930:Leucine; D014633:Valine; D011188:Potassium