Na+-independent transport of bipolar and cationic amino acids across the luminal membrane of the small intestine

1997 ◽  
Vol 272 (4) ◽  
pp. R1060-R1068 ◽  
Author(s):  
B. G. Munck ◽  
L. K. Munck

The role of sodium in transport of bipolar and cationic amino acids and their interactions were examined in vitro by measuring unidirectional influx across the brush-border membrane of intact rat jejunal and rabbit ileal epithelia. The chloride-dependent and beta-alanine inhibitable B(0,+) present in rabbit ileum was blocked by combining inhibition by beta-alanine with Na(+)- or Cl(-)-free conditions. Under these conditions, lysine influx across the brush-border membrane is Na+ independent. All Na+-independent influx of cationic and bipolar amino acids is by a system b(0,+) equivalent in the brush-border membrane of both species, where a system y+ is not present. System b(0,+) is shown to be a potent exchanger of intracellular leucine for extracellular lysine and of intracellular lysine for extracellular leucine. The model used to explain leucine stimulation of mucosa to serosa lysine transport can explain Na+ dependence of net lysine absorption. On the assumption that b(0,+) in situ, like the transporter induced by retroperitoneal brown adipose tissue in Xenopus laevi oocytes, acts as an obligatory exchanger, this model can also explain the effects of lysine on short-circuit current and net transport of sodium and the effect on transport capacity by preincubation at Na+-free conditions.

1986 ◽  
Vol 251 (1) ◽  
pp. F125-F131
Author(s):  
R. W. Chesney ◽  
N. Gusowski ◽  
M. Padilla ◽  
S. Lippincott

Alterations in the intake of sulfur amino acids (SAA) changes the rat renal brush-border membrane uptake of the beta-amino acid, taurine. A low-SAA diet enhances and a high-taurine diet reduces uptake (Chesney et al., Kidney Int. 24: 588-594, 1983). Neither the low-SAA diet nor the high-taurine diet alters the time course or concentration-dependent accumulation of the sulfur amino acids methionine and cystine or of inorganic sulfate. By contrast the uptake of beta-alanine, another beta-amino acid that competes with taurine, is greater in animals on the low-SAA diet. The high-taurine diet does not change beta-alanine uptake. The plasma levels of taurine are altered by dietary change, but not the values for methionine and cystine. This study indicates that renal adaptation is expressed for beta-alanine, a nonsulfur-containing beta-amino acid. By contrast, methionine, cystine, and sulfate, which participate in a variety of synthetic and conjugative processes, are not conserved by the renal brush-border surface following ingestion of either a low-methionine and -cystine diet or high-taurine diet.


1992 ◽  
Vol 262 (4) ◽  
pp. G609-G615 ◽  
Author(s):  
L. K. Munck ◽  
B. G. Munck

This study describes the influx of taurine and beta-alanine across the brush-border membrane of rabbit distal ileum. The kinetics of JmcTau [concentration at which half-maximal activation occurs (K1/2) = 41 microM and Jmax = 24 nmol.cm-2.h-1] are consistent with the kinetics of taurine uptake by jejunal brush-border vesicles. The taurine carrier differs from the beta-alanine carrier by being insensitive to leucine inhibition and by the jejunoileal variation of influx along the small intestine. The K1/2 for sodium and chloride activation of the beta-alanine carrier (48 and 8 mM, respectively) differ markedly from the values reported for the taurine carrier. In addition, taurine is not transported by the beta-alanine carrier. Thus the study demonstrates that the taurine and beta-alanine carriers are separate entities, and it adds to the imino and the taurine carriers the beta-alanine carrier as a third chloride-dependent intestinal transporter of amino acids.


2002 ◽  
Vol 205 (6) ◽  
pp. 745-758 ◽  
Author(s):  
W. S. Marshall ◽  
J. A. Howard ◽  
R. R. F. Cozzi ◽  
E. M. Lynch

SUMMARYSections of posterior intestine of the euryhaline killifish Fundulus heteroclitus adapted to sea water were stimulated by the calcium ionophore ionomycin (1 μmol l–1) in combination with agents to elevate intracellular cyclic AMP levels, 0.5 mmol l–1 dibutyryl-cyclic AMP (db-cAMP) with 0.1 mmol l–1 3-isobutyl-1-methylxanthine (IBMX). Intestinal bag preparations from recently fed animals (but not from overnight unfed animals) changed from fluid absorption (+18.9±8.30 μl cm–2 h–1 , N=8) in the untreated control period to net fluid secretion after stimulation (–7.43±1.30 μl cm–2 h–1, N=8, P<0.01; means ± s.e.m.), indicative of the capacity of teleost intestine to undergo secretion. Posterior intestinal pieces mounted in vitro in Ussing-style membrane chambers showed net Cl– uptake (+2.245±0.633 μequiv cm–2 h–1, N=7) that turned to net secretion following stimulation by ionomycin + db-cAMP + IBMX (–3.809±1.22 μequiv cm–2 h–1, N=7, P<0.01). Mucosal application of the anion channel blocker 1 mmol l–1 diphenylamine-2-carboxylate (DPC) after ionomycin + db-cAMP + IBMX treatment significantly reduced serosal-to-mucosal unidirectional Cl– flux (P<0.001), net Cl– flux (P<0.05), short-circuit current (Isc, P<0.001) and tissue conductance (Gt, P<0.001), while 0.1 mmol l–1 4,4′-diisothiocyano-2,2′-stilbene-disulphonic acid (DIDS, a blocker of anion exchange) was without effect. Stimulation by db-cAMP + IBMX (no ionomycin) significantly increased unidirectional fluxes, Isc and Gt but did not produce net Cl– secretion. Ionomycin alone produced a transient increase in Isc but had no effect on Gt and caused no significant changes in unidirectional or net Cl– fluxes. Addition of db-cAMP + IBMX after ionomycin treatment produced net secretion of Cl– and large increases in unidirectional fluxes and Gt. Cystic fibrosis transmembrane conductance regulator (CFTR) was immunocytochemically localized with a monoclonal mouse antibody to the carboxy terminus and found to be present in the cytoplasm and basolateral membranes of all enterocytes and in the brush-border membrane of some cells, whereas NKCC immunofluorescence, demonstrating the presence of the Na+/K+/2Cl– cotransporter, was present in the cytoplasm and brush-border membrane. We conclude that the teleost intestine is capable of salt and fluid secretion only if intracellular Ca2+ and cyclic AMP pathways are stimulated together and that this secretion appears to involve activation of CFTR ion channels in the apical membrane of a subpopulation of enterocytes.


1979 ◽  
Vol 236 (5) ◽  
pp. E534
Author(s):  
J J Hajjar ◽  
D M Murphy ◽  
R L Scheig

The mechanism of inhibition of alanine absorption by Na ricinoleate has been examined in the rabbit intestine. This fatty acid in a concentration of 2--5 mM inhibits alanine absorption in vivo and in vitro. The inhibition is more evident in the jejunum than in the ileum. Strips of ileal mucosa treated with Na ricinoleate gain Na. Sodium ricinoleate inhibits alanine influx across rabbit ileum, even in the presence of a sodium gradient across these cells. The results suggest that the main action of Na ricinoleate is on the alanine-transport system at the brush-border membrane. The fatty acid may also inhibit amino acid absorption by increasing intestinal cell Na concentration, which results in a decreased Na gradient across the brush-border membrane.


1970 ◽  
Vol 56 (5) ◽  
pp. 621-639 ◽  
Author(s):  
Stanley G. Schultz ◽  
Lorna Yu-Tu ◽  
Osvaldo O. Alvarez ◽  
Peter F. Curran

Glutamate and aspartate influxes across the brush border of rabbit intestine are saturable processes that are subject to competitive inhibition and are markedly influenced by the Na concentration in the mucosal solution. Lowering the Na concentration increases the amino acid concentration needed to elicit a half-maximal influx but does not significantly affect the maximal influx. The interaction between Na and anionic amino acid influx can be described by the same kinetic model that has been applied to the influxes of neutral amino acids and lysine. Comparison of the kinetic parameters for anionic, neutral, and cationic amino acids suggests that amino acid charge influences (a) the stability of the binary (amino acid-site) complex and (b) the affinity of this binary complex for the subsequent binding of Na. A mechanistic interpretation of these interactions is proposed.


1994 ◽  
Vol 189 (1) ◽  
pp. 55-67
Author(s):  
R Parthasarathy ◽  
W R Harvey

The time-dependent fluorescence intensity of an intravesicular potential-sensitive dye was used to probe the real-time kinetics of potential difference (PD)-dependent amino acid/Na+ symport at pH9 into brush-border membrane vesicles obtained from larval Manduca sexta midgut. Neutral amino acids (alanine, proline) are symported at higher rates as the vesicles are hyperpolarized. The symport rates of acidic (glutamate) and basic (arginine) amino acids are almost PD-independent. The half-saturation constant of alanine is PD-independent between -108 and -78 mV, although the maximal symport velocity increases by half as the voltage is increased. Amino acid throughput is evidently enhanced as the relatively high transmembrane PDs (> 150 mV, lumen positive) measured in vivo are approached. The half-saturation concentrations of Na+ were in the range 15-40 mmol l-1 for most of the amino acids examined and increased with voltage for alanine. The Vmax observed as a function of cation or amino acid concentration increased as the vesicle was hyperpolarized in the case of leucine and alanine. The data support the hypothesis that carrier and substrates are at equilibrium inasmuch as substrate translocation seems to be the rate-determining step of symport.


2019 ◽  
Vol 20 (6) ◽  
pp. 1504 ◽  
Author(s):  
Subha Arthur ◽  
Palanikumar Manoharan ◽  
Shanmuga Sundaram ◽  
M Rahman ◽  
Balasubramanian Palaniappan ◽  
...  

Na-amino acid co-transporters (NaAAcT) are uniquely affected in rabbit intestinal villus cell brush border membrane (BBM) during chronic intestinal inflammation. Specifically, Na-alanine co-transport (ASCT1) is inhibited secondary to a reduction in the affinity of the co-transporter for alanine, whereas Na-glutamine co-transport (B0AT1) is inhibited secondary to a reduction in BBM co-transporter numbers. During chronic intestinal inflammation, there is abundant production of the potent oxidant peroxynitrite (OONO). However, whether OONO mediates the unique alteration in NaAAcT in intestinal epithelial cells during chronic intestinal inflammation is unknown. In this study, ASCT1 and B0AT1 were inhibited by OONO in vitro. The mechanism of inhibition of ASCT1 by OONO was secondary to a reduction in the affinity of the co-transporter for alanine, and secondary to a reduction in the number of co-transporters for B0AT1, which were further confirmed by Western blot analyses. In conclusion, peroxynitrite inhibited both BBM ASCT1 and B0AT1 in intestinal epithelial cells but by different mechanisms. These alterations in the villus cells are similar to those seen in the rabbit model of chronic enteritis. Therefore, this study indicates that peroxynitrite may mediate the inhibition of ASCT1 and B0AT1 during inflammation, when OONO levels are known to be elevated in the mucosa.


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