Ionic circuit analysis of K+/H+ antiport and amino acid/K+ symport energized by a proton-motive force in Manduca sexta larval midgut vesicles.

1994 ◽  
Vol 196 (1) ◽  
pp. 77-92
Author(s):  
F G Martin ◽  
W R Harvey
Keyword(s):  
Amino Acid ◽  
Membrane Conductance ◽  
Ionic Concentration ◽  
Circuit Analysis ◽  
Metabolic Energy ◽  
Transport Systems ◽  
Mathematical Treatment ◽  
Acid Uptake ◽  
State Equations ◽  
Transport Pathways

Amino acid/K+ symport (cotransport) across a model epithelium, the lepidopteran midgut, is energized by an electrogenic H+ V-ATPase (H+ pump) in parallel with an electrophoretic K+/H+ antiporter (exchanger). Attempts to analyze this process using well-known equilibrium thermodynamic equations (Nernst, Gibbs), diffusion equations (Nernst, Planck, Einstein, Goldman, Hodgkin, Katz) and equations based on Ohm's law (Hodgkin, Huxley) have all encountered major difficulties. Although they are useful for analyzing nerve/muscle action potentials, these state equations assume that brief perturbations in membrane conductance, gm, and membrane voltage, Vm, occur so rapidly that no other parameters are significantly disturbed. However, transport studies often extend for minutes, even for hours. Perturbation of one parameter in complex transport systems invariably results in a state change as all of the other elements adjust to the prolonged stress. The development of a comprehensive mathematical treatment for transport systems that contain pumps and porters (transporters) has been hampered by the empirical nature of the concept of membrane permeability and conductance. The empirical definition of permeability was developed before pumps and porters were known. Thus, 'permeability' is a gross parameter that, in practice if not in theory, could describe all transport pathways including pumps, porters and channels. To surmount these difficulties, we have applied ionic circuit analysis to vesicular systems containing insect midgut transport proteins. In this analysis, pumps, porters and channels, as well as ionic concentration gradients and membrane capacitance, are components of ionic circuits that function to transform metabolic energy (e.g. from ATP hydrolysis) into useful metabolic work (e.g. amino acid uptake). Computer-generated by an H+ V-ATPase to K+/2H+ antiport and amino acid/K+ symport in the lepidopteran midgut.

Placental toxicology: tobacco smoke, abused drugs, multiple chemical interactions, and placental function

1991 ◽  
Vol 3 (4) ◽  
pp. 355 ◽  
Author(s):  
BV Sastry
Keyword(s):  
Amino Acid ◽  
Amino Acid Transport ◽  
Drugs Of Abuse ◽  
Amino Acid Uptake ◽  
Transport Systems ◽  
Acid Uptake ◽  
Acid Transport ◽  
Ach Release ◽  
Placental Blood ◽  
Abused Drugs

There are increasing numbers of reports on the tobacco smoking and ingestion of abused drugs (e.g. morphine, cocaine) by pregnant women and the effects of the substances on the developing fetus and newborn infant. The passage of drugs and chemicals from the mother to the fetus is influenced by the placental transport and metabolism of the substances. Further, these drugs and chemicals affect the nutrient transport systems in the placenta. The three major drugs of abuse-nicotine, morphine and cocaine-depress both active amino-acid uptake by human placental villi and transplacental amino-acid transport by reason of the drugs' influence on placental cholinergic and opiate systems. Part of this depression (10-16%) is not reversible. Nicotine blocks the cholinergic receptor and thus blocks acetylcholine (ACh)-facilitated amino-acid transport. Morphine stimulates opiate kappa receptors and depresses ACh release. Cocaine blocks Ca2+ influx and thus blocks ACh release. ACh causes dilation of blood vessels and maintains placental blood flow by the activation of endothelial muscarinic receptors. By interfering with ACh release and placental blood flow, the three drugs of abuse may depress the diffusion of amino acids and other nutrients from the trophoblast into the placental circulation. Three regulatory systems are delineated for amino-acid uptake by the placenta: placental ACh, phospholipid N-methyltransferase, and the gammaglutamyl cycle. These systems operate in concert with one another and are dependent on cellular formation of adenosine 5'-triphosphate (ATP). Placental hypoxia induced by carbon monoxide and other tobacco gases depresses the energy-dependent processes and thus the ATP levels of placental cells. Maternal tobacco smoking and drug abuse cause placental insufficiencies for amino-acid transport, which may partially explain the fetal intrauterine growth retardation caused by these substances. Part of the amino-acid deficits may be compensated for by the induction of new amino-acid transport systems. Specific receptors or drug-binding proteins for the three drugs of abuse are present in the placenta. A DNA adduct selective for maternal smoking has been demonstrated in the placenta. DNA adducts selective for cocaine, morphine and other environmental chemicals have yet to be demonstrated ins the placenta.

Amino acid uptake systems in Bacteroides ruminicola

1979 ◽  
Vol 25 (10) ◽  
pp. 1161-1168 ◽  
Author(s):  
Roselynn M. W. Stevenson
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Nitrogen Sources ◽  
Amino Acid Uptake ◽  
Defined Medium ◽  
High Rate ◽  
Transport Systems ◽  
Acid Uptake ◽  
Chemical Structures ◽  
Kinetic Inhibition

Uptake of amino acids by Bacteroides ruminicola was observed in cells grown in a complete defined medium, containing ammonia as the nitrogen source. A high rate of uptake occurred only in fresh medium, as an inhibitory substance, possibly acetate, apparently accumulated during growth. All amino acids except proline were taken up and incorporated into cold trichloroacetic acid precipitable material. Different patterns of incorporation and different responses to 2,4-dinitrophenol and potassium ferricyanide indicated multiple uptake systems were involved. Kinetic inhibition patterns suggested six distinct systems were present for amino acid uptake, with specificities related to the chemical structures of the amino acids. Thus, the failure of free amino acids to act as sole nitrogen sources for growth of B. ruminicola is not due to the absence of transport systems for these compounds.

Myocardial amino acid transport by canine sarcolemma vesicles

1987 ◽  
Vol 252 (6) ◽  
pp. H1070-H1076
Author(s):  
L. H. Young ◽  
B. L. Zaret ◽  
E. J. Barrett
Keyword(s):  
Amino Acid ◽  
Sarcoplasmic Reticulum ◽  
Membrane Vesicles ◽  
Amino Acid Uptake ◽  
Ventricular Myocardium ◽  
Transport Systems ◽  
Acid Uptake ◽  
Leucine Uptake ◽  
Alanine Transport ◽  
Canine Ventricular Myocardium

The transport of L-alanine and L-leucine into membrane vesicles isolated from mature canine ventricular myocardium was studied. Transport was assessed in purified sarcolemma and in vesicles differentially enriched either for sarcolemma or sarcoplasmic reticulum to further localize these transport systems. An imposed inward gradient of a NaNO3 stimulated uptake of L-alanine but not L-leucine by these vesicles. Amino acid uptake by these vesicles occurred into an osmotically active space. The stimulatory effect of Na+ on alanine transport was most striking in the purified sarcolemma vesicles, where Na+-stimulated alanine flux was 45 +/- 14 pmol X mg-1 X min-1. Furthermore, Na+-dependent alanine transport activity appeared to copurify with Na+-K+-ATPase activity, which served as a marker for sarcolemma membrane when these activities were compared in the three different membrane preparations. Leucine transport by sarcolemma was not altered by an imposed Na+ gradient. However, leucine uptake was a saturable function of extravesicular leucine and was inhibited by valine. In contrast, in sarcoplasmic reticulum membrane vesicles leucine uptake increased proportionately with increasing media leucine and was unaffected by valine. Our results demonstrate the feasibility of directly studying the transport of naturally occurring amino acids in membrane vesicles from mammalian heart, and the presence of Na+-dependent alanine transport system and a Na+-independent leucine transporter in the sarcolemma but not in sarcoplasmic reticulum of canine ventricular myocardium.

In vivo muscle amino acid transport involves two distinct processes

2004 ◽  
Vol 287 (1) ◽  
pp. E136-E141 ◽  
Author(s):  
Sharon Miller ◽  
David Chinkes ◽  
David A. MacLean ◽  
Dennis Gore ◽  
Robert R. Wolfe
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Transport ◽  
Interstitial Fluid ◽  
Muscle Blood Flow ◽  
Amino Acid Uptake ◽  
Transport Systems ◽  
Acid Uptake ◽  
Acid Transport ◽  
Rate Limiting

We have tested the hypothesis that transit through the interstitial fluid, rather than across cell membranes, is rate limiting for amino acid uptake from blood into muscle in human subjects. To quantify muscle transmembrane transport of naturally occurring amino acids, we developed a novel 4-pool model that distinguishes between the interstitial and intracellular fluid compartments. Transport kinetics of phenylalanine, leucine, lysine, and alanine were quantified using tracers labeled with stable isotopes. The results indicate that interstitial fluid is a functional compartment insofar as amino acid kinetics are concerned. In the case of leucine and alanine, transit between blood and interstitial fluid was potentially rate limiting for muscle amino acid uptake and release in the postabsorptive state. For example, in the case of leucine, the rate of transport between blood and interstitial fluid compared with the corresponding rate between interstitial fluid and muscle was 247 ± 36 vs. 610 ± 95 nmol·min−1·100 ml leg−1, respectively ( P < 0.05). Our results are consistent with the process of diffusion governing transit from blood to interstitial fluid without selectivity, and of specific amino acid transport systems with varying degrees of efficiency governing transit from interstitial fluid to muscle. These results imply that changes in factors that affect the transit of amino acids from blood through interstitial fluid, such as muscle blood flow or edema, could play a major role in controlling the rate of muscle amino acid uptake.

ASC system activity is altered by development of cell polarity in trophoblast from human placenta

1993 ◽  
Vol 265 (1) ◽  
pp. C212-C217 ◽  
Author(s):  
T. C. Furesz ◽  
C. H. Smith ◽  
A. J. Moe
Keyword(s):  
Amino Acid ◽  
Primary Cultures ◽  
Amino Acid Uptake ◽  
Selective Inhibition ◽  
Isobutyric Acid ◽  
Neutral Amino Acid ◽  
Transport Systems ◽  
Bewo Cell ◽  
Acid Uptake

Pathways of neutral amino acid uptake were investigated in vitro during differentiation of primary cultures of trophoblast isolated from full-term human placentas and a clone (b30) of the BeWo cell line. Inhibition of initial alanine (0.1 microM) uptake by 2-(methylamino)isobutyric acid and unlabeled alanine revealed two Na(+)-dependent systems and one Na(+)-independent transporter. Characterization of these transporters, by selective inhibition, suggested system A, ASC, and L-like transporters. Concomitant with formation of microvillous membrane and syncytium, system ASC activity decreased from 16.1 +/- 2.8 pmol.mg DNA-1.min-1 at 24 h to 2.4 +/- 1.1 pmol.mg DNA-1.min-1 at 72 h. Na(+)-independent alanine uptake increased from 6.0 +/- 2.0 to 12.9 +/- 0.9 pmol.mg DNA-1.min-1 at 24 and 72 h, respectively. Similarly, alpha-(methylamino)isobutyric acid-insensitive, Na(+)-dependent activity in b30 cells (100 microM alanine) decreased from 6.5 +/- 1.6 to 1.2 +/- 1.2 nmol.mg DNA-1.min-1 for control and forskolin-treated cells, respectively. We conclude that membrane specialization accompanying fusion and differentiation of the cytotrophoblast to form syncytiotrophoblast results in a polarization of neutral amino acid transport systems.

Neutral amino acid transport systems of microvillous membrane of human placenta

1988 ◽  
Vol 254 (6) ◽  
pp. C773-C780 ◽  
Author(s):  
L. W. Johnson ◽  
C. H. Smith
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Transport ◽  
Plasma Membranes ◽  
Competitive Inhibition ◽  
Transport Systems ◽  
Acid Uptake ◽  
Acid Transport ◽  
Independent System ◽  
High Affinity

Placental transport produces concentrations of amino acids in fetal blood greater than those of maternal blood. Competitive inhibition studies of zwitterionic amino acid transport in isolated vesicles from the microvillous (maternal facing) plasma membranes of syncytiotrophoblast defined three transport systems: 1) a sodium-dependent system that supports methylaminoisobutyric acid (MeAIB) transport and has the characteristics of an A system; 2) a sodium-independent system with a high affinity for leucine and other amino acids with branched or aromatic side chains; and 3) a sodium-independent system with a preference for alanine as a substrate. The two sodium-independent systems could be further discriminated by marked specificity for trans stimulation with alanine or with leucine. System ASC, known to be present in whole placenta, and the neutral brush-border or imino systems of other polarized epithelia were apparently absent. Kinetic characteristics of the A system make it the probable primary driving force for concentrative transfer of its substrate amino acids to the fetus. Characteristics of the high-affinity leucine system demonstrated that it is saturated by normal serum leucine concentrations. Regulation of either system has the potential to alter placental amino acid uptake and transfer to the fetus.

scholarly journals Evidence that cycloleucine affects the high-affinity systems of amino acid uptake in cultured human fibroblasts

1984 ◽  
Vol 224 (1) ◽  
pp. 309-315 ◽  
Author(s):  
M Feneant ◽  
N Moatti ◽  
J Maccario ◽  
M Gautier ◽  
S Guerroui ◽  
...  
Keyword(s):  
Amino Acid ◽  
Transport System ◽  
Human Fibroblasts ◽  
Diffusion Constant ◽  
Amino Acid Uptake ◽  
Cell Uptake ◽  
Transport Systems ◽  
Acid Uptake ◽  
High Affinity ◽  
Cultured Human Fibroblasts

The influence of cycloleucine on kinetic parameters of uptake of L-alanine, L-proline and L-leucine into cultured human fibroblasts was examined under initial-rate conditions with substrate concentrations of 0.05-10 mM and 5 mM-cycloleucine. Kinetic data obtained by computer analysis showed that, in the absence of cycloleucine, cell uptake was heterogeneous for each amino acid. L-Alanine and L-leucine entered by two transport systems with different affinities; L-proline was taken up by one saturable transport system plus a diffusion-like process. This heterogeneity disappeared in the presence of cycloleucine, since the high-affinity systems were no longer detectable. The remaining process had the same kinetic constants as the low-affinity system for alanine and leucine and a KD similar to the diffusion constant for proline. The influence of cycloleucine on the amino acid uptake was not specific either to the amino acid concerned or to a particular transport system, since the three neutral amino acid-transport systems, A, ASC and L, were involved in these experiments. This influence was shown to be unaffected by the absence of Na+ (for leucine uptake). ATP content of the cells was identical in the presence or in the absence of cycloleucine.

scholarly journals A novel pathway of nutrient absorption in crustaceans: branchial amino acid uptake in the green shore crab ( Carcinus maenas )

2017 ◽  
Vol 284 (1868) ◽  
pp. 20171298 ◽  
Author(s):  
Tamzin A. Blewett ◽  
Greg G. Goss
Keyword(s):  
Amino Acid ◽  
Carcinus Maenas ◽  
High Capacity ◽  
Amino Acid Uptake ◽  
Shore Crab ◽  
Acid Uptake ◽  
Transport Pathways ◽  
Amino Acid Transport System ◽  
High Affinity

Estuaries are environments enriched with dissolved nutrients such as amino acids. To date, marine arthropods are the only invertebrate group that have not been demonstrated to access this potentially important nutrient resource. Using in vitro gill perfusion techniques, we sought to investigate the ability of the green shore crab ( Carcinus maenas ) to take up the amino acid l -leucine directly from the water. Investigation of the concentration-dependent transport kinetics of radiolabelled l -leucine showed that there are two specific transport pathways across Carcinus gills, one with high affinity and low capacity, and the other with high capacity and low affinity. Using putative competitive substrates and reduced sodium preparations, we were able to identify the putative amino acid transport system associated with high-affinity uptake. This is the first study to demonstrate the absorption of dissolved organic nutrients across the gill epithelium of a marine arthropod.

Mutations in Saccharomyces cerevisiae which confer resistance to several amino acid analogs

1990 ◽  
Vol 10 (6) ◽  
pp. 2941-2949
Author(s):  
J H McCusker ◽  
J E Haber
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Amino Acids ◽  
Amino Acid ◽  
Nitrogen Source ◽  
Transport Systems ◽  
Acid Uptake ◽  
Acid Analog ◽  
Amino Acid Analogs ◽  
Amino Acid Analog ◽  
Complementation Groups

Four new complementation groups of mutations which confer resistance to several amino acid analogs in Saccharomyces cerevisiae are described. These mutants were isolated on medium containing urea as the nitrogen source, in contrast to previous studies that had used medium containing proline. All four resistance to amino acid analog (raa) complementation groups appear to confer resistance by reducing amino acid analog and amino acid uptake. In some genetic backgrounds, raa leu2 and raa thr4 double mutants are inviable, even on rich medium. The raa4 mutation may affect multiple amino acid transport systems, since raa4 mutants are unable to use proline as a nitrogen source. raa4 is, however, unlinked to a previously described amino acid analog resistance and proline uptake mutant, aap1, or to the general amino acid permease mutant gap1. Both raa4 and gap1 prevent uptake of [3H]leucine in liquid cultures. The raa1, raa2, and raa3 mutants affect only a subset of the amino acid analogs and amino acids affected by raa4. The phenotypes of raa1, -2, and -3 mutants are readily observed on agar plates but are not seen in uptake and incorporation of amino acids measured in liquid media.

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