In the small intestine, cationic amino acids are transported by y(+)-like and b(0,+)-like systems present in the luminal side of the epithelium. Here, we report the characterization of a b(0,+)-like system in the apical membrane of the chicken jejunum, and its properties as an amino acid exchanger. Analysis of the brush border membrane by Western blot points out the presence of rBAT (protein related to b0,+ amino acid transport system) in these membranes. A functional mechanism for amino acid exchange across this system was established by kinetic analysis measuring fluxes at varying substrate concentrations both in internal (in) and external (out) vesicle compartments. This intestinal b(0,+)-like system functions for L-arginine as an obligatory exchanger since its transport capacity increases 100-200 fold in exchange conditions, thus suggesting an important role in the intestinal absorption of cationic amino acids. The kinetic analysis of Argin efflux velocities is compatible with the formation of a ternary complex and excludes a model involving a ping-pong mechanism. The binding affinity of Argout is higher than that of Argin, suggesting a possible order of binding (Argout first) for the formation of the ternary complex during the exchange cycle. A model of double translocation pathways with alternating access is discussed.

The properties of l-lysine transport in chicken jejunum have been studied in brush border membrane vesicles isolated from 6-wk-old birds. l-lysine uptake was found to occur within an osmotically active space with significant binding to the membrane. The vesicles can accumulate l-lysine against a concentration gradient, by a membrane potential-sensitive mechanism. The kinetics of l-lysine transport were described by two saturable processes: first, a high affinity-transport system (KmA = 2.4 +/- 0.7 micromol/L) which recognizes cationic and also neutral amino acids with similar affinity in the presence or absence of Na+ (l-methionine inhibition constant KiA, NaSCN = 21.0 +/- 8.7 micromol/L and KSCN = 55.0 +/- 8. 4 micromol/L); second, a low-affinity transport mechanism (KmB = 164. 0 +/- 13.0 micromol/L) which also recognizes neutral amino acids. This latter system shows a higher affinity in the presence of Na+ (KiB for L-methionine, NaSCN = 1.7 +/- 0.3 and KSCN = 3.4 +/- 0.9 mmol/L). L-lysine influx was significantly reduced with N-ethylmaleimide (0.5 mmol/L) treatment. Accelerative exchange of extravesicular labeled l-lysine was demonstrated in vesicles preloaded with 1 mmol/L l-lysine, l-arginine or l-methionine. Results support the view that l-lysine is transported in the chicken jejunum by two transport systems, A and B, with properties similar to those described for systems b0,+ and y+, respectively.

The influx ofl-lysine into apical vesicles from the chicken jejunum occurs through two systems, one with low Michaelis constant ( K m) and features of system b0,+ and the other with relatively high K m forl-lysine and with properties of system y+. In the present study the effect of a lysine-enriched diet (Lys, containing 68 g l-lysine/kg dietary protein, control animals 48 g/kg) onl-lysine uptake through both transport systems was investigated. Results show that 1) lysine enrichment had no effect on either body weight or the efficiency of food utilization. 2) In Lys-fed animals, the mediatedl-lysine influx was best fitted to the two-system model with y+and b0,+ activity. 3) In the presence of an Na+ gradient, totall-lysine uptake is significantly higher in Lys-fed animals than in control birds (about 40% increase). 4) Lys diet increases K mb0,+6-fold (KSCN gradient) and 12-fold (NaSCN gradient) and maximum velocity ( V max) by 6- and 20-fold, respectively. The effects of Lys enrichment on the y+-like system are only observed on the V max and in the presence of a Na+ gradient (30% increase). 5) Na+ is involved in the activation of the transport process in the Lys-fed chickens, but there is no correlation between external Na+concentration and l-lysine influx. In conclusion, both b0,+- and y+-like transport systems are upregulated by dietary lysine but with different kinetic profiles; the high-capacity y+-like carrier shows a V maxincrease without changes in K m, whereas the low-capacity b0,+-like system shows an increase in V max as well as in the K m.

The intestinal transport of L‐methionine has been investigated in brush‐border membrane vesicles isolated from the jejunum of 6‐week‐old chickens. L‐Methionine influx is mediated by passive diffusion and by Na+‐dependent and Na+‐independent carrier‐mediated mechanisms. In the absence of Na+, cis‐inhibition experiments with neutral and cationic amino acids indicate that two transport components are involved in L‐methionine influx: one sensitive to L‐lysine and the other sensitive to 2‐aminobicyclo[2.2.1]heptane‐2‐carboxylic acid (BCH). The L‐lysine‐sensitive flux is strongly inhibited by L‐phenylalanine and can be broken down into two pathways, one sensitive to N‐ethylmaleimide (NEM) and the other to L‐glutamine and L‐cystine. The kinetics of L‐methionine influx in Na+‐free conditions is described by a model involving three transport systems, here called a,b and c: systems a and b are able to interact with cationic amino acids but differ in their kinetic characteristics (system a: Km= 2.2 ± 0.3 μM and Vmax= 0.13 ± 0.005 pmol (mg protein)−1 (2 s)−1; system b: Km= 3.0 ± 0.3 mM and Vmax= 465 ± 4.3 pmol (mg protein)−1 (2 s)−1); system c is specific for neutral amino acids, has a Km of 1.29 ± 0.08 mM and a Vmax of 229 ± 5.0 pmol (mg protein)−1 (2 s)−1 and is sensitive to BCH inhibition. The Na+‐dependent component can be inhibited by BCH and L‐phenylalanine but cannot interact either with cationic amino acids or with α‐(methylamino)isobutyrate (MeAIB). The kinetic analysis of L‐methionine influx under a Na+ gradient confirms the activity of the above described transport systems a and b. System a is not affected by the presence of Na+ while system b shows a 3‐fold decrease in the Michaelis constant and a 1.4‐fold increase in Vmax. In the presence of Na+, the BCH‐sensitive component can be subdivided into two pathways: one corresponds to system c and the other is Na+ dependent and has a Km of 0.64 ± 0.013 mM and a Vmax of 391 ± 2.3 pmol (mg protein)−1 (2 s)−1. It is concluded that L‐methionine is transported in the chicken jejunum by four transport systems, one with functional characteristics similar to those of system bo, + (system a); a second (system b) similar to system y+, which we suggest naming y+m to account for its high Vmax for L‐methionine transport in the absence of Na+; a third (system c) which is Na+ independent and has similar properties to system L; and a fourth showing Na+ dependence and tentatively identified with system B.

The methionine hydroxy analogue DL-2-hydroxy-4-(methylthio)butanoic acid (HMB) is commonly used as a supplemental source of methionine in commercial animal diets. The HMB free acid is an aqueous solution that contains 88% product in an equilibrium mixture of monomer, dimer, and polymeric compounds. The present study examines whether the presence of these nonmonomeric forms reduces the absorption of the hydroxy analogue in the chicken small intestine. In vivo and in vitro methodologies were used to compare the intestinal absorption of an HMB product containing mainly monomer (HMB-PCM) with commercial HMB. The results from the in vivo perfusion of the jejunum showed no significant differences between the 2 hydroxy analogue sources in monomer absorption from the intestinal lumen, tissue accumulation, or plasma concentration. The results also indicate that the nonmonomeric forms are hydrolyzed during perfusion. Moreover, monomer tissue accumulation in everted sacs showed no significant differences between substrates, either in the presence or in the absence of a H+-gradient; a higher value was observed in the jejunum and ileum in comparison with the duodenum. Similarly, serosal appearance in H+-gradient conditions did not differ significantly between substrates, and it showed the same regional profile as in tissue accumulation. Oligomer hydrolysis was confirmed in vitro without significant differences between segments. In conclusion, the presence of nonmonomeric forms is not a limiting factor in HMB absorption, apparently because of the hydrolytic capacity of intestinal mucosa, as confirmed by experiments in vivo and in vitro.