monensin and 3-3--4--5-tetrachlorosalicylanilide

Peptostreptococcus anaerobius converted glutamine stoichiometrically to ammonia and pyroglutamic acid, and the Eadie-Hofstee plot of glutamine transport was biphasic. High-affinity, sodium-dependent glutamine transport (affinity constant [Kt] of 1.5 microM) could be driven by the chemical gradient of sodium, and more than 20 mM sodium was required for half-maximal velocity. High-affinity glutamine transport was not stimulated or inhibited by a membrane potential (delta psi). Low-affinity glutamine transport had a rate which was directly proportional to the external glutamine concentration, required less than 100 microM sodium, and was inhibited strongly by a delta psi. Cells which were treated with N,N-dicyclohexylcarbodiimide to inhibit the F1F0 ATPase still generated a delta psi but did so only if the external glutamine concentration was greater than 15 mM. Low-affinity glutamine uptake could not be saturated by as much as 200 mM glutamine, but glutamine-1 accounts for only a small fraction of the total glutamine at physiological pH values (pH 6 to 7). On the basis of these results, it appeared that the low-affinity glutamine transport was an electrogenic mechanism which was converting a chemical gradient of glutamine-1 into a delta psi. Other mechanisms of delta psi generation (electrogenic glutamine-pyroglutamate or -ammonium exchange) could not be demonstrated.

Topics: Biological Transport; Carbon Radioisotopes; Dicyclohexylcarbodiimide; Glutamine; Kinetics; Membrane Potentials; Monensin; Peptostreptococcus; Potassium; Salicylanilides; Sodium; Sodium Chloride; Time Factors; Tromethamine

Mixed ruminal bacteria (80 mg N/liter) degraded casein and soluble soy protein rapidly (.68 and .72 mg N/[liter.min], respectively), but ammonia was produced at a slower rate (.08 and .10 mg N/[liter.min], respectively). Because there was little increase in cell protein, ammonia production could not account for all the degraded protein. Large quantities of non-ammonia, non-protein nitrogen (NAN-NPN) accumulated, and this NAN-NPN reacted more strongly (2- to 14-fold) with ninhydrin after it was treated with 6 N HCl (110 degrees C, 24 h) or pronase E. Even after 96 h of incubation, 10% of the protein N was still found in the NAN-NPN pool. Monensin had little effect on protein degradation, but it caused a large decrease in ammonia production (P less than .05) and an increase in NAN-NPN (P less than .05). These results indicated that significant quantities of peptide N could not be degraded by ruminal microorganisms and that monensin could increase peptide flow from the rumen. Because 3,3',4',5-tetrachlorosalicylanide, a protonophore that inhibits both Gram-positive and Gram-negative bacteria, did not cause a greater decrease (P greater than .05) in ammonia than monensin, an ionophore that is primarily effective against Gram-positive bacteria, it seemed that the "protein sparing" of monensin could largely be explained by its inhibition of Gram-positive bacteria.

Topics: Ammonia; Animals; Bacteria; Caseins; Cattle; Deamination; Female; Glycine max; Monensin; Peptides; Plant Proteins, Dietary; Proteins; Proton-Translocating ATPases; Rumen; Salicylanilides; Soybean Proteins