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PubMed1982

Stimulation of Na+ -dependent amino acid uptake by activation of the Ca2+ -dependent K+ channel in the Ehrlich ascites tumor cell.

The activation of Ca2+ -dependent K+ channel by propranolol or by ascorbate-phenazine methosulphate stimulates Na+ -dependent transport of alpha-aminoisobutyric acid. This stimulation arises from a membrane hyperpolarization due to the specific increase of membrane K+ conductance. The same treatment does not modify the Na+ -independent uptake of the norbornane amino acid.

Amino Acids↗

PubMed1990

Placental transfer of glucose and amino acids in intrauterine growth retardation: studies with substrate analogs in the awake guinea pig.

After experimentally inducing growth retardation by unilateral uterine artery ligation in midpregnancy, placental blood flow (PBF) (microsphere technique) and placental transfer of 14C-aminoisobutyric acid (AIB) and 3H-methylglucose (MG) were studied at d 44, 50, or 63 of gestation in 19 chronically catheterized awake guinea pigs. At d 44, fetal wt (FW) and placental wt were reduced by 16 and 18%, respectively, in the ligated horn and FW/PBF was increased by 122%. Placental efficiency to transfer MG and AIB (fetal dpm/g placenta) was maintained in the ligated horn, feto-placental extraction (feto-placental dpm/PBF) was increased, and placental transfer of the substrate analogs (fetal dpm/g fetus) was unchanged. At d 50 and 63, placental wt and FW were reduced in proportion (approximately 40%) in the growth-retarded group and FW/PBF was increased by 80 and 51%, respectively. Placental transfer of AIB was reduced by 33% at d 50 and by 18% at d 63. In addition, placental efficiency to transfer AIB was reduced by 36% at d 50 and by 22% at d 63 in the growth-retarded group. Fetal uptake of MG per g fetus was slightly reduced (-6%) at d 50 of gestation, but unaffected at d 63. Extraction of MG from the maternal PBF was increased, whereas the wt-specific transfer capacity of the placenta was unaltered.(ABSTRACT TRUNCATED AT 250 WORDS)

3-O-Methylglucose↗

PubMed1982

Antigen-specific stimulation of amino acid transport in bovine lymphocytes.

Treatment of bovine lymphocytes isolated from animals which were either infected with Mycobacterium bovis or sensitized to a purified protein derivative (PPD-B) from this organism induced an increase in the transport of alpha-aminoisobutyric acid (AIB) and alpha-methylaminoisobutyric acid (MeAIB). PPD-B did not stimulate these transport activities in lymphocytes from nonsensitized animals. The transport stimulation was first measurable after about 7 hours of treatment, reached about a two-fold enhancement after 20 hours, and continued to increase to 30- to 40-fold after 6 days. The stimulation of AIB transport was inhibited by both ouabain and cycloheximide. Experiments to determine transport system specificities in nonstimulated lymphocytes showed that MeAIB transport was primarily by the Na+-dependent, A-system, and leucine transport was mostly by Na+-independent system(s). In contrast, AIB transport was about 25% by the A-system, 25% by at least one Na+-dependent, non-A-system, and 50% by one or more Na+-independent system(s). Analysis of the three components of AIB transport after treatment with PPD-B showed that: 1) transport by both the A-system and the Na+-independent system(s) was stimulated; 2) A-system transport was stimulated to a larger extent than Na+-independent transport; and 3) Na+-dependent, non-A-system transport was not stimulated significantly.

Amino Acids↗

PubMed1975

Plasma amino acid levels and development of hepatic gluconeogenesis in the newborn rat.

The metabolism of endogenous and exogenous amino acids has been characterized during a 16-h fast after birth in the rat. Eighteen of 22 amino acids showed a decrease in plasma concentration up to 16 h, the most profound and sustained changes affecting those quantitatively important in gluconeogenesis. The hepatic accumulation of injected [14C]aminoisobutyric acid showed a progressive rise after birth. The in vivo conversion of 14C-labeled lactate, alanine, serine, and glutamine to [14C]glucose increased for 6 h, but all except glutamine showed a decline by 16 h. The in vitro conversion of several gluconeogenic substrates (10mM), however, increased with time in each instance. These data confirm that the capacity for hepatic gluconeogenesis and maintenance of blood glucose concentration appears immediately after birth. Nevertheless, profound hypoglycemia recurs at 16 h and responds only minimally and transiently to exogenous gluconeogenic substrate loads. In contrast, the fed newborn maintains normoglycemia, higher endogenous amino acid levels, and the capacity for substrate conversion at this time. The mechanism for stimulation of hepatic gluconeogenic pathways thus is present in both fasted and fed neonatal rats. However, owing to insufficient energy sources to sustain gluconeogenesis and to inadequate gluconeogenic substrate, the rat is unable to maintain normoglycemia if fasted 16 h.

Acetoacetates↗

PubMed1980

Regulation of amino acid transport in chicken embryo fibroblasts by purified multiplication-stimulating activity (MSA).

Enhanced amino acid transport is observed when quiescent cultures of chicken embryo fibroblasts are stimulated to proliferate by the addition of purified multiplication-stimulating activity (MSA). This increase in amino acid transport is an early event occurring prior to the onset of DNA synthesis in stimulated cells. Results indicate that the changes in transport activity, as measured by alpha-aminoisobutyric acid (AIB) uptake, are due to stimulation of only the Na+-dependent A transport system. There is little or no change in the activities of transport systems ASC, L, or Ly+ upon exposure to MSA. A kinetic analysis shows this increased activity is due to a change in Vmax while Km remains unaltered. Continuous exposure to the stimulus is required to maintain the increased level of transport activity and the presence of inhibitors of RNA and protein synthesis significantly inhibits the response. Results also indicate that a similar specific increase in the A transport system is initiated when RSV tsNY68 infected cells are shifted to the permissive temperature. It appears that the A system of mediation is emerging as a strategic regulatory site for cell function.

Amino Acids↗

PubMed1993

Regulation of amino acid transport and protein metabolism in myotubes derived from chicken muscle satellite cells by insulin-like growth factor-I.

The effects of insulin and insulin-like growth factor-I (IGF-I) on amino acid transport and protein metabolism were compared in myotubes derived from chicken breast muscle satellite cells. Protein synthesis was assessed by continuous labelling with [3H]-tyrosine. Protein degradation was estimated by the release of trichloroacetic acid (TCA) soluble radioactivity by cells which had been previously labelled with [3H]-tyrosine for 3 days. Amino acid transport was measured in myotubes incubated in Dulbecco's modified Eagle's medium (DMEM) 0.5% bovine serum albumin (BSA) with or without insulin or IGF-I. Subsequent [3H]-aminoisobutyric acid (AIB) uptake was then measured in amino acid-free medium. IGF-I was more efficient than insulin at equimolar concentration (3.2 nmol/l) in stimulating protein synthesis (127 and 113% of basal, respectively) and inhibiting protein degradation (32% and 13% inhibition of protein degradation following 4 h incubation). Half maximal effective concentrations for stimulation of AIB uptake were 0.27 +/- 0.03 nmol/l and 34.8 +/- 3.1 nmol/l for IGF-I and insulin respectively, with maximal stimulation of about 340% of basal. Cycloheximide (3.6 mumol/l) diminished IGF-I-stimulated AIB uptake by 55%. Chicken growth hormone had no effect on basal AIB uptake in these cells and neither glucagon nor dexamethasone had an effect on basal or IGF-I-stimulated AIB uptake. This study demonstrates an anabolic effect for IGF-I in myotubes derived from primary chicken satellite cells which is mediated by the type I IGF receptor, since the cation-independent mannose 6-phosphate receptor does not bind IGF-II in chicken cells.

Amino Acids↗

PubMed1998

Na+ cotransport by metabolizable and nonmetabolizable amino acids stimulates a glucose-regulated insulin-secretory response.

The involvement of Na+ in insulin-secretory responses to metabolizable and nonmetabolizable amino acids known to be cotransported with Na+, were examined using islet-derived BRIN-BD11 cells. At stimulatory (16.7 mM) glucose, 10 mM of l-alanine, alpha-aminoisobutyric acid (AIB) or l-proline stimulated 1.3- to 10. 4-fold (p < 0.01) insulin-secretory responses. In each case, these effects were significantly greater than those observed at nonstimulatory (1.1 mM) glucose (p < 0.01). While, tetrodotoxin blockade of voltage-dependent Na+ channels exerted no significant effect on insulin release, Na/K pump blockade with ouabain significantly promoted the amino acid-induced effects (p < 0.05). Replacement of extracellular Na+ with equimolar N-methyl-d-glucamine+ and omission of extracellular K+ or Ca2+ were all effective in removing the actions of each amino acid, confirming the critical role of ionic fluxes in the secretory responses to these amino acids. Collectively these results demonstrate that metabolizable and nonmetabolizable amino acids can induce glucose-dependent insulin-secretory responses by modulating electrogenic Na+ transport.

Alanine↗

PubMed1985

Effects of ethanol ingestion on amino acid uptake in the dog liver in vivo.

The effect of ethanol ingestion on the uptake of labeled amino acids was studied in the in situ autoperfused dog liver. Ethanol was administered orally, as a 15% water solution, in a dose of 4 g/kg body weight/day as the only source of water for 2 days. Amino acid uptake was measured in anesthetized dogs by means of the single-passage, multiple-tracer dilution technique. In control animals, hepatic uptake of 14C-glycine, 3H-alpha-aminoisobutyric acid (3H-AIB) and 3H-L-leucine were 50, 15 and 66%, respectively. In the ethanol-treated dogs, glycine and AIB uptake was reduced by 70 and 63%, respectively. L-leucine uptake was reduced by only 23%. The plasma concentration of the naturally occurring amino acids was significantly increased after ethanol treatment, probably due to a reduced influx into hepatocytes. Simultaneous measurements of hemodynamic parameters showed a significant increase in the portal vein pressure of ethanol-treated animals, whereas the portal vein blood flow and hepatic extracellular volume were unaffected.

Amino Acids↗

PubMed1994

Purification, characterization and inhibition of dihydropyrimidinase from rat liver.

Dihydropyrimidinase (DHPase) was purified 564-fold over the initial rat liver extract, using heat, ammonium sulfate fractionation, DEAE-Sepharose CL-6B, carboxymethyl-Sepharose CL-6B, hydroxyapatite and Sephacryl S-300 chromatography. The purified enzyme was shown to be homogeneous by gel electrophoresis both in the presence and absence of SDS. Its molecular mass, determined by gel filtration, was 215 kDa and the subunit mass was 54 kDa. DHPase catalyzed the reversible cyclization of 5,6-dihydrouracil (H2Ura) to N-carbamoyl-beta-alanine or 5,6-dihydrothymine (H2Thy) to N-carbamoyl-beta-aminoisobutyric acid. Authentic 5-bromo-5,6-dihydrouracil (BrH2Ura) and commercially available H2Thy were racemic. However, these 5-substituted 5,6-dihydropyrimidines were hydrolyzed by over 96% and 98%, respectively, by DHPase. These results suggest that dihydropyrimidinase has no stereo specificities for 5-substituents of H2Ura. The addition of H2Ura and H2Thy competitively inhibited the enzyme activity against BrH2Ura. However, the addition of N-carbamoyl-beta-alanine or N-carbamoyl-beta-amino-isobutyric acid showed hyperbolic mixed-type inhibition, when BrH2Ura was used as the substrate. The values of the dissociation constants of BrH2Ura, N-carbamoyl-beta-alanine and N-carbamoyl-beta-aminoisobutyric acid were 17 microM, 0.38 mM and 0.38 mM, respectively. DHPase from the rat liver contains 4 mol Zn2+/mol active enzyme, presumably one atom/subunit. Zn2+ also inhibited the hydrolysis of BrH2Ura by the enzyme. The Ki for Zn2+ as an inhibitor of DHPase was 23 microM, and the maximum rate of inactivation was 0.057 min-1 at 37 degrees C. H2Ura and H2Thy protected the enzyme activity from Zn2+ inactivation.

Amidohydrolases↗

PubMed1989

Peculiarities of amino acid transport in Schizosaccharomyces pombe: effects of growth medium.

Transport systems for amino acids in the wild-type strain of Schizosaccharomyces pombe are not constitutive. During growth on different media no transport of acidic, neutral and basic amino acids is detectable. To acquire the ability to transport amino acids, cells must be preincubated with a metabolic source of energy, such as glucose. The appearance of transport activity is associated with protein synthesis (suppression by cycloheximide) at all phases of culture growth. After such preincubation the initial rate of amino acid uptake depends on the phase of growth of the culture and on the amount of glucose in the growth medium but not on the nitrogen source used. L-Proline and 2-aminoisobutyric acid are practically not transported under any of the conditions tested.

Amino Acids↗

PubMedDate not supplied

Dinaline inhibits amino acid transport and proliferation of colon carcinoma cells in vitro.

The human colon carcinoma cell line SW707 was incubated with different concentrations (7 microM - 540 microM) of the novel antineoplastic drug 4-amino-N(2'-aminophenyl)benzamide (GOE1734, dinaline) to study the effects on 3H-alpha-aminoisobutyric acid (AIB) - and 14C-methionine uptake as well as on 3H-thymidine incorporation into DNA. Additionally, adenine nucleotide pools were determined by high performance liquid chromatography (h.p.l.c.), and cell cycle distribution by flow cytometry. 24 h exposure to low concentrations of dinaline caused mild cell proliferation which turned into a cytostatic effect within a further 24 h without drug exposure. High concentrations of dinaline, however, were found to be cytostatic and then cytotoxic after corresponding time intervals. Dinaline caused a concentration-dependent decrease in sodium-dependent AIB uptake. Immediately after exposure the effect ranged from 67%-36% of control uptake. One day later an incomplete recovery not exceeding 70% of control was found for the three highest concentrations. This differed significantly from sodium-dependent methionine uptake which was initially decreased by 24%-36% but recovered within one hour and was enhanced one day after exposure. Significant differences were also measured for sodium-independent AIB uptake and sodium-independent methionine uptake immediately, 1 and 4 h after exposure. Adenine nucleotide pools were mildly increased immediately and 4 h after exposure. Thymidine incorporation into DNA was decreased by 70% to 84% immediately after exposure, and no full recovery was seen in the subsequent observation period. Cell cycle analyses revealed a block at the S phase entrance. Changes in thymidine incorporation and S phase fraction were highly correlated. The inhibition of the sodium-dependent uptake of AIB might indicate interaction of dinaline with cell membrane functions at low concentrations, the increase in methionine uptake points to enhanced protein synthesis following drug withdrawal and the impaired thymidine incorporation into DNA results from the cytostatic effect of dinaline. Taken together, the results show that SW707 colon carcinoma cells exposed to dinaline demonstrate distinct but reversible changes in amino acid transport, protein metabolism, DNA synthesis and cell proliferation, thus giving a correlation with observations in vivo.

Adenine Nucleotides↗

PubMed1991

Metabolic actions of insulin-like growth factor I in cultured glomerular mesangial cells.

Glomerular mesangial cells in culture have been reported to possess a considerable number of receptors specific to insulin-like growth factor I (IGF-I), with very small number of receptors specific to insulin. To explore acute metabolic effects of IGF-I on mesangial cells, uptake of glucose and amino acid was measured in the presence of IGF-I or insulin. IGF-I stimulated D-[U-14C]glucose incorporation, 2-deoxy[1-3H]glucose uptake and alpha-[methyl-3H]aminoisobutyric acid (AIB) uptake into cultured mesangial cells by 139.8% +/- 2.1%, 116.6% +/- 1.7%, and 214.9% +/- 12.8% (percent of basal), respectively. Similar maximal stimulation was also induced by insulin, while the ED50 of IGF-I to stimulate these uptake systems (9.98 +/- 2.36, 3.45 +/- 1.86, and 3.35 +/- 0.40 ng/mL, respectively) was significantly lower than that of insulin (120.8 +/- 28.5, 61.8 +/- 7.7, and 76.3 +/- 17.5, respectively). These results indicate that, in cultured glomerular mesangial cells, IGF-I induces acute metabolic effects, possibly through its own receptors.

Amino Acids↗

PubMed1996

Effect of epidermal growth factor on placental amino acid transport and regulation of epidermal growth factor receptor expression of hepatocyte in rat.

We investigated the effect of epidermal growth factor (EGF) on amino acid transport in the rat placenta and the hormonal and nutritional regulation of hepatic expression of epidermal growth factor receptor (EGFR). Fetuses born to maternal rats administered EGF were larger than control fetuses, while fetuses born to mothers administered EGF antibody were smaller than controls. The fetomaternal amino acid concentration ratio (fetal blood/maternal blood) was higher in the EGF-treated group than in the control group, and was lower in the EGF antibody-treated group. EGF treatment stimulated 14C-aminoisobutyric acid, isoleucine and alanine uptake by placental explants as did treatment with insulin-like growth factor-1 and insulin, which have been reported to stimulate the active amino acid transport. Thus, EGF promoted amino acid transport in the rat placenta and influenced fetal growth as a result. In addition, retinol induced a 170% increase of EGF binding to rat hepatocytes and insulin induced a 120% increase of EGF binding, while amino acids (isoleucine and serine) had no effect on EGF binding. This result indicated that there is hormonal regulation of EGF binding to hepatocytes and that changes of some hormone levels may affect hepatic EGFR expression.

Alanine↗

PubMedDate not supplied

[Probable genetic independence of four hereditary characteristics peculiar to Mongoloid populations].

Several hereditary characteristics are found in Mongoloid populations (Asian and Amerindian) with a much higher frequency than in other populations. These are the mongoloid facial features, the presence of alcohol dehydrogenase beta 2, the presence of inactive mitochondrial aldehyde dehydrogenase 2, and the high rate of urinary excretion of beta aminoisobutyric acid. Analyses carried out in France and Switzerland have been made to research a possible correlation between these diverse characteristics. This correlation does not seem to exist. With regard to the excretion of beta aminoisobutyric acid, the hypothetical role of mercury in dental fillings has been checked in Caucasoid subjects. If this role is not completely excluded, it is far from being statistically significant.

Adolescent↗

PubMed1972

Increased uptake of amino acids and 2-deoxy-D-glucose by virus-transformed cells in culture.

Transformed and nontransformed cells in tissue culture differ in their rate of uptake of certain nutrients, as determined by a polyester-coverslip technique. A 2.5- to 3.5-fold increased rate of uptake of alpha-aminoisobutyric acid, cycloleucine, and 2-deoxy-D-glucose was observed with polyoma virus-transformed baby hamster kidney (BHK) 21 cells and simian virus 40 (SV40)-transformed BALB/3T3 (mouse fibroblast) cells, compared to their nontransformed counterparts. Kinetic analysis suggested that the increased uptake by cells transformed with virus was associated with a 3-fold greater V(max), with no detectable changes in apparent K(m). Limited studies also revealed increased initial rates of uptake by murine sarcoma virus-transformed rat liver cells, as compared to the parental line. Exposure of cells to concanavalin A and wheat-germ agglutinin led to significant reductions in amino-acid uptake by both transformed and nontransformed cells; however, transformed cells showed a greater decrease in uptake after exposure to wheat-germ agglutinin. Increased initial rates of uptake of certain amino acids and sugars may be a feature common to transformed cells, compared to their parental control.

Amino Acids↗

PubMed1975

A protein from polymorphonuclear leukocytes (LEM) which affects the rate of hepatic amino acid transport and synthesis of acute-phase globulins.

A proteinaceous secretion from phagocytizing polymorphonuclear leukocytes, termed "leukocytic endogenous mediator" (LEM), has been shown to have marked effects on hepatic amino acid transport and RNA and protein synthesis. A single injection of LEM results in a marked accumulation of labeled nonmetabolizable model amino acids in the liver of normal rats. The LEM-stimulated uptake of amino acids by liver was observed in adrenalectomized, hypophysectomized, thyroidectomized, or diabetic rats and could not be duplicated by pharmacological doses of a large variety of hormones. In addition, LEM stimulated an increased uptake of alpha-aminoisobutyric acid by isolated livers during their perfusion in vitro. LEM also stimulated an increased incorporation of orotic acid into hepatic RNA of intact rats, especially into the bound ribosomal fraction. This increased synthesis of RNA preceded an enhanced hepatic production of a number of the acute-phase plasma globulins. LEM did not stimulate the adenylate cyclase-cAMP system in liver and was not found to utilize this system as a second messenger. Thus, the effects of LEM in stimulating hepatic amino acid transport appear to be direct, without mediation by other hormones, and to be independent of cAMP. On the other hand, the ability of LEM to stimulate RNA and acute phase globulin synthesis in liver may require the presence of physiological quantities of hormones such as adrenal corticoids.

Adenylyl Cyclases↗

PubMed1990

Effect of exercise and obesity on skeletal muscle amino acid uptake.

The genetically obese Zucker rat has a reduced capacity to deposit dietary protein in skeletal muscle. To determine whether amino acid uptake by muscle of obese Zucker rats is impaired, soleus strip (SOL) and epitrochlearis (EPI) muscles from 10-wk-old lean and obese Zucker rats were studied in vitro by use of [14C]alpha-aminoisobutyric acid (AIB). Muscles from fasted rats were incubated under basal conditions at rest or after a 1-h treadmill run at 8% grade. To equate total work completed, lean and obese rats ran at 27 and 20 m/min, respectively. Muscles were pinned at resting length, preincubated for 30 min at 37 degrees C in Krebs-Ringer bicarbonate buffer containing 5 mM glucose under 95% O2-5% CO2, and then incubated up to 3 h in Krebs-Ringer bicarbonate with 0.5 mM AIB, [14C]AIB, and [3H]inulin as a marker of extracellular fluid. Basal AIB uptake in EPI and SOL from obese rats was significantly reduced by 40 and 30% (P less than 0.01), respectively, compared with lean rats. For both lean and obese rats, exercise increased (P less than 0.05) basal AIB uptake in EPI and SOL, but the relative increases were greater in the obese rats (EPI 54% and SOL 71% vs. EPI 32% and SOL 37%). These results demonstrate that genetically obese Zucker rats have reduced basal skeletal muscle amino acid uptake and suggest that physical inactivity may partially contribute to this defect.

Amino Acids↗

PubMed2007

Bombesin analogs containing alpha-amino-isobutyric acid with potent anticancer activity.

Six octapeptide bombesin (BN) analogs were synthesized by substituting alpha-aminoisobutyric acid (Aib), in place of Ala9 or Gly11, or both, in the [D-Phe6, desMet14]-BN (6-14) sequence: D-Phe6-Gln7-Trp8-Ala9-Val10-Gly11-His12-Leu13-NH2 (P0). Additionally, Leu13 was replaced with isoleucine in two analogs and one of the analogs was butanoylated at the N-terminus. The antiproliferative activity of the analogs was tested in vitro on human pancreatic (MiaPaCa-2) and colon cancer (SW620, HT29 and PTC) cell lines using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The analogs demonstrated anticancer activity in the above cell lines at concentrations ranging from 0.01 nM to 1 microM. One of the analogs, P6, was evaluated for in vivo tumor regression in a xenograft model of human primary colon cancer in athymic nude mice and was found to cause significant reduction in tumor volume. NMR and molecular dynamics (MD) simulation studies for this analog revealed the presence of a mixed 3(10)/alpha-helical structure. This study demonstrates that the designed BN analogs retain their anticancer activity after the incorporation of the constrained amino acid, Aib, and are potential molecules for future use in cancer therapy and drug targeting.

Amino Acid Sequence↗