scholarly journals CAT2-mediated l-arginine transport and nitric oxide production in activated macrophages

1999 ◽  
Vol 340 (2) ◽  
pp. 549-553 ◽  
Author(s):  
Donald K. KAKUDA ◽  
Matthew J. SWEET ◽  
Carol L. MACLEOD ◽  
David A. HUME ◽  
Daniel MARKOVICH
Keyword(s):  
Amino Acid ◽  
Transport System ◽  
Transport Systems ◽  
No Production ◽  
Activated Macrophages ◽  
Amino Acid Transport System ◽  
Cationic Amino Acid ◽  
Arginine Transport ◽  
Arginine Uptake ◽  
Expression Studies

Activated macrophages require L-arginine uptake to sustain NO synthesis. Several transport systems could mediate this L-arginine influx. Using competition analysis and gene-expression studies, amino acid transport system y+ was identified as the major carrier responsible for this activity. To identify which of the four known y+ transport-system genes is involved in macrophage-induced L-arginine uptake, we used a hybrid-depletion study in Xenopus oocytes. Cationic amino acid transporter (CAT) 2 antisense oligodeoxyribonucleotides abolished the activated-macrophage-mRNA-induced L-arginine transport. Together with expression studies documenting that CAT2 mRNA and protein levels are elevated with increased L-arginine uptake, our data demonstrate that CAT2 mediates the L-arginine transport that is required for the raised NO production in activated J774 macrophages.

Characterization of l-arginine transport in adrenal cells: effect of ACTH

2006 ◽  
Vol 291 (2) ◽  
pp. E291-E297 ◽  
Author(s):  
Esteban M. Repetto ◽  
Vanesa Pannunzio ◽  
Francisco Astort ◽  
Camila Martinez Calejman ◽  
Marcos Besio Moreno ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Amino Acid ◽  
Transport System ◽  
Zona Fasciculata ◽  
Acid Uptake ◽  
Uptake System ◽  
Amino Acid Transport System ◽  
Cationic Amino Acid ◽  
Arginine Transport

Nitric oxide synthesis depends on the availability of its precursor l-arginine, which could be regulated by the presence of a specific uptake system. In the present report, the characterization of the l-arginine transport system in mouse adrenal Y1 cells was performed. l-arginine transport was mediated by the cationic/neutral amino acid transport system y+L and the cationic amino acid transporter (CAT) y+ in Y1 cells. These Na+-independent transporters were identified by their selectivity for neutral amino acids in both the presence and absence of Na+ and by the effect of N-ethylmaleimide. Transport data correlated to expression of genes encoding for CAT-1, CAT-2, CD-98, and y+LAT-2. A similar expression profile was detected in rat adrenal zona fasciculata. In addition, cationic amino acid uptake in Y1 cells was upregulated by ACTH and/or cAMP with a concomitant increase in nitric oxide (NO) production.

scholarly journals Arginine Homeostasis in J774.1 Macrophages in the Context of Mycobacterium bovis BCG Infection

2006 ◽  
Vol 188 (13) ◽  
pp. 4830-4840 ◽  
Author(s):  
Meliza T. Talaue ◽  
Vishwanath Venketaraman ◽  
Manzour Hernando Hazbón ◽  
Marcy Peteroy-Kelly ◽  
Anjali Seth ◽  
...  
Keyword(s):  
Amino Acid ◽  
Mycobacterium Bovis ◽  
Amino Acid Transport ◽  
Host Cells ◽  
Transport Systems ◽  
Acid Transport ◽  
Intracellular Growth ◽  
Cationic Amino Acid ◽  
Arginine Transport ◽  
Arginine Uptake

ABSTRACT The competition for l-arginine between the inducible nitric oxide synthase and arginase contributes to the outcome of several parasitic and bacterial infections. The acquisition of l-arginine, however, is important not only for the host cells but also for the intracellular pathogen. In this study we observe that strain AS-1, the Mycobacterium bovis BCG strain lacking the Rv0522 gene, which encodes an arginine permease, perturbs l-arginine metabolism in J774.1 murine macrophages. Infection with AS-1, but not with wild-type BCG, induced l-arginine uptake in J774.1 cells. This increase in l-arginine uptake was independent of activation with gamma interferon plus lipopolysaccharide and correlated with increased expression of the MCAT1 and MCAT2 cationic amino acid transport genes. AS-1 infection also enhanced arginase activity in resting J774.1 cells. Survival studies revealed that AS-1 survived better than BCG within resting J774.1 cells. Intracellular growth of AS-1 was further enhanced by inhibiting arginase and ornithine decarboxylase activities in J774.1 cells using l-norvaline and difluoromethylornithine treatment, respectively. These results suggest that the arginine-related activities of J774.1 macrophages are affected by the arginine transport capacity of the infecting BCG strain. The loss of Rv0522 gene-encoded arginine transport may have induced other cationic amino acid transport systems during intracellular growth of AS-1, allowing better survival within resting macrophages.

scholarly journals Increased l-arginine uptake and inducible nitric oxide synthase activity in aortas of rats with heart failure

2001 ◽  
Vol 280 (2) ◽  
pp. H859-H867 ◽  
Author(s):  
Peter B. Stathopulos ◽  
Xiangru Lu ◽  
Ji Shen ◽  
Jeremy A. Scott ◽  
James R. Hammond ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Heart Failure ◽  
Amino Acid ◽  
Nitric Oxide Synthase ◽  
No Production ◽  
Cationic Amino Acid ◽  
Arginine Transport ◽  
Arginine Uptake ◽  
Oxide Synthase

l-Arginine crosses the cell membrane primarily through the system y+ transporter. The aim of this study was to investigate the role of l-arginine transport in nitric oxide (NO) production in aortas of rats with heart failure induced by myocardial infarction. Tumor necrosis factor-α levels in aortas of rats with heart failure were six times higher than in sham rats ( P < 0.01). l-Arginine uptake was increased in aortas of rats with heart failure compared with sham rats ( P < 0.01). Cationic amino acid transporter-2B and inducible (i) nitric oxide synthase (NOS) expression were increased in aortas of rats with heart failure compared with sham rats ( P < 0.05). Aortic strips from rats with heart failure treated with l-arginine but not d-arginine increased NO production ( P < 0.05). The effect ofl-arginine on NO production was blocked byl-lysine, a basic amino acid that shares the same system y+ transporter withl-arginine, and by the NOS inhibitor N G-nitro-l-arginine methyl ester (l-NAME). Treatment with l-lysine andl-NAME in vivo decreased plasma nitrate and nitrite levels in rats with heart failure ( P < 0.05). Our data demonstrate that NO production is dependent on iNOS activity andl-arginine uptake and suggest that l-arginine transport plays an important role in enhanced NO production in heart failure.

Diminished L-arginine bioavailability in hypertension

2004 ◽  
Vol 107 (4) ◽  
pp. 391-397 ◽  
Author(s):  
Monique B. MOSS ◽  
Tatiana M. C. BRUNINI ◽  
Roberto SOARES de MOURA ◽  
Lúcia E. NOVAES MALAGRIS ◽  
Norman B. ROBERTS ◽  
...  
Keyword(s):  
Amino Acid ◽  
Blood Cells ◽  
Human Platelets ◽  
Transport Systems ◽  
No Production ◽  
Hypertensive Patients ◽  
Spontaneously Hypertensive ◽  
Cationic Amino Acid ◽  
Arginine Transport ◽  
Amino Acid Profiles

L-Arginine is the precursor of NO (nitric oxide), a key endogenous mediator involved in endothelium-dependent vascular relaxation and platelet function. Although the concentration of intracellular L-arginine is well above the Km for NO synthesis, in many cells and pathological conditions the transport of L-arginine is essential for NO production (L-arginine paradox). The present study was designed to investigate the modulation of L-arginine/NO pathway in systemic arterial hypertension. Transport of L-arginine into RBCs (red blood cells) and platelets, NOS (NO synthase) activity and amino acid profiles in plasma were analysed in hypertensive patients and in an animal model of hypertension. Influx of L-arginine into RBCs was mediated by the cationic amino acid transport systems y+ and y+L, whereas, in platelets, influx was mediated only via system y+L. Chromatographic analyses revealed higher plasma levels of L-arginine in hypertensive patients (175±19 μmol/l) compared with control subjects (137±8 μmol/l). L-Arginine transport via system y+L, but not y+, was significantly reduced in RBCs from hypertensive patients (60±7 μmol·l−1·cells−1·h−1; n=16) compared with controls (90±17 μmol·l−1·cells−1·h−1; n=18). In human platelets, the Vmax for L-arginine transport via system y+L was 86±17 pmol·109 cells−1·min−1 in controls compared with 36±9 pmol·109 cells−1·min−1 in hypertensive patients (n=10; P<0.05). Basal NOS activity was decreased in platelets from hypertensive patients (0.12±0.02 pmol/108 cells; n=8) compared with controls (0.22±0.01 pmol/108 cells; n=8; P<0.05). Studies with spontaneously hypertensive rats demonstrated that transport of L-arginine via system y+L was also inhibited in RBCs. Our findings provide the first evidence that hypertension is associated with an inhibition of L-arginine transport via system y+L in both humans and animals, with reduced availability of L-arginine limiting NO synthesis in blood cells.

Estradiol augments while progesterone inhibits arginine transport in human endothelial cells through modulation of cationic amino acid transporter-1

2015 ◽  
Vol 309 (4) ◽  
pp. R421-R427 ◽  
Author(s):  
Ohad S. Bentur ◽  
Doron Schwartz ◽  
Tamara Chernichovski ◽  
Merav Ingbir ◽  
Talia Weinstein ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Amino Acid ◽  
Protein Content ◽  
Amino Acid Transporter ◽  
Female Rats ◽  
Cationic Amino Acid Transporter ◽  
Cationic Amino Acid ◽  
Arginine Transport ◽  
Arginine Uptake ◽  
Acid Transporter

Decreased generation of nitric oxide (NO) by endothelial NO synthase (eNOS) characterizes endothelial dysfunction (ECD). Delivery of arginine to eNOS by cationic amino acid transporter-1 (CAT-1) was shown to modulate eNOS activity. We found in female rats, but not in males, that CAT-1 activity is preserved with age and in chronic renal failure, two experimental models of ECD. In contrast, during pregnancy CAT-1 is inhibited. We hypothesize that female sex hormones regulate arginine transport. Arginine uptake in human umbilical vein endothelial cells (HUVEC) was determined following incubation with either 17β-estradiol (E2) or progesterone. Exposure to E2 (50 and 100 nM) for 30 min resulted in a significant increase in arginine transport and reduction in phosphorylated CAT-1 (the inactive form) protein content. This was coupled with a decrease in phosphorylated MAPK/extracellular signal-regulated kinase (ERK) 1/2. Progesterone (1 and 100 pM for 30 min) attenuated arginine uptake and increased phosphorylated CAT-1, phosphorylated protein kinase Cα (PKCα), and phosphorylated ERK1/2 protein content. GO-6976 (PKCα inhibitor) prevented the progesterone-induced decrease in arginine transport. Coincubation with both progesterone and estrogen for 30 min resulted in attenuated arginine transport. While estradiol increases arginine transport and CAT-1 activity through modulation of constitutive signaling transduction pathways involving ERK, progesterone inhibits arginine transport and CAT-1 via both PKCα and ERK1/2 phosphorylation, an effect that predominates over estradiol.

Modulation of the l-arginine/nitric oxide signalling pathway in vascular endothelial cells

2004 ◽  
Vol 71 ◽  
pp. 143-156 ◽  
Author(s):  
Amanda W. Wyatt ◽  
Joern R. Steinert ◽  
Giovanni E. Mann
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Amino Acid ◽  
Protein Kinase ◽  
Signalling Pathway ◽  
No Production ◽  
Dependent Manner ◽  
Amino Acid Transport System ◽  
Membrane Hyperpolarization ◽  
Cationic Amino Acid

Nitric oxide (NO) is synthesized from l-arginine, and in endothelial cells influx of l-arginine is mediated predominantly via Na+-independent cationic amino acid transporters. Constitutive, Ca2+-calmodulin-sensitive eNOS (endothelial nitric oxide synthase) metabolizes l-arginine to NO and l-citrulline. eNOS is present in membrane caveolae and the cytosol and requires tetrahydrobiopterin, NADPH, FAD and FMN as additional cofactors for its activity. Supply of l-arginine for NO synthesis appears to be derived from a membrane-associated compartment distinct from the bulk intracellular amino acid pool, e.g. near invaginations of the plasma membrane referred to as 'lipid rafts' or caveolae. Co-localization of eNOS and the cationic amino acid transport system y+ in caveolae in part explains the 'arginine paradox', related to the phenomenon that in certain disease states eNOS requires an extracellular supply of l-arginine despite having sufficient intracellular l-arginine concentrations. Vasoactive agonists normally elevate [Ca2+]i (intracellular calcium concentration) in endothelial cells, thus stimulating NO production, whereas fluid shear stress, 17ϐ-oestradiol and insulin cause phosphorylation of the serine/threonine protein kinase Akt/protein kinase B in a phosphoinositide 3-kinase-dependent manner and activation of eNOS at basal [Ca2+]i levels. Adenosine causes an acute activation of p42/p44 mitogen-activated protein kinase and NO release, with membrane hyperpolarization leading to increased system y+ activity in fetal endothelial cells. In addition to acute stimulatory actions of D-glucose and insulin on l-arginine transport and NO synthesis, gestational diabetes, intrauterine growth retardation and pre-eclampsia induce phenotypic changes in the fetal vasculature, resulting in alterations in the l-arginine/NO signalling pathway and regulation of [Ca2+]i. These alterations may have significant implications for long-term programming of the fetal cardiovascular system.

scholarly journals Development and polarization of cationic amino acid transporters and regulators in the human placenta

2000 ◽  
Vol 278 (6) ◽  
pp. C1162-C1171 ◽  
Author(s):  
Paul T.-Y. Ayuk ◽  
Colin P. Sibley ◽  
Paul Donnai ◽  
Stephen D'Souza ◽  
Jocelyn D. Glazier
Keyword(s):  
Amino Acid ◽  
Membrane Vesicles ◽  
First Trimester ◽  
Transport Systems ◽  
Amino Acid Transporters ◽  
Rt Pcr ◽  
Plasma Membrane Vesicles ◽  
Michaelis Constant ◽  
Cationic Amino Acid ◽  
Arginine Transport

We have investigatedl-arginine transport systems in the human placental syncytiotrophoblast across gestation using purified microvillous (MVM) and basal (BM) plasma membrane vesicles. In MVM from first-trimester and term placentas, l-arginine transport was by systems y+ and y+L. In BM (term placentas), however, there was evidence for system y+L only. The Michaelis constant of system y+L was significantly lower ( P< 0.05) in first-trimester compared with term MVM and lower in term MVM compared with BM ( P < 0.05). There was no functional evidence for system b0+ in term MVM or BM. Cationic amino acid transporter (CAT) 1, CAT 4, and 4F2hc were detected using RT-PCR in placentas throughout gestation. rBAT was not detected in term placentas. An ∼85-kDa and an ∼135-kDa protein was detected by Western blotting in MVM under reducing and nonreducing conditions, respectively, consistent with the 4F2hc monomer and the 4F2hc-light chain dimer, and their expression was significantly higher ( P< 0.05) in term compared with first-trimester MVM. These proteins were not detected in BM despite functional evidence for system y+L. These data suggest different roles for 4F2hc in the development and polarization of cationic amino acid transporters in the syncytiotrophoblast.

Cytoskeletal regulation of the l-arginine/NO pathway in pulmonary artery endothelial cells

2001 ◽  
Vol 280 (3) ◽  
pp. L465-L473 ◽  
Author(s):  
Sergey I. Zharikov ◽  
Alla A. Sigova ◽  
Sifeng Chen ◽  
Michael R. Bubb ◽  
Edward R. Block
Keyword(s):  
Endothelial Cells ◽  
Pulmonary Artery ◽  
Actin Cytoskeleton ◽  
Actin Polymerization ◽  
The State ◽  
No Production ◽  
Actin Microfilaments ◽  
Cationic Amino Acid ◽  
Arginine Transport ◽  
Arginine Uptake

We investigated possible involvement of the actin cytoskeleton in the regulation of thel-arginine/nitric oxide (NO) pathway in pulmonary artery endothelial cells (PAEC). We exposed cultured PAEC to swinholide A (Swinh), which severs actin microfilaments, or jasplakinolide (Jasp), which stabilizes actin filaments and promotes actin polymerization, or both. After treatment, the state of the actin cytoskeleton,l-arginine uptake mediated by the cationic amino acid transporter-1 (CAT-1), Ca2+/calmodulin-dependent (endothelial) NO synthase (eNOS) activity and content, and NO production were examined. Jasp (50–100 nM, 2 h treatment) induced a reversible activation ofl-[3H]arginine uptake by PAEC, whereas Swinh (10–50 nM) decreased l-[3H]arginine uptake. The two drugs could abrogate the effect of each other onl-[3H]arginine uptake. The effects of both drugs on l-[3H]arginine transport were not related to changes in expression of CAT-1 transporters. Swinh (50 nM, 2 h) and Jasp (100 nM, 2 h) did not change eNOS activities and contents in PAEC. Detection of NO in PAEC by the fluorescent probe 4,5-diaminofluorescein diacetate showed that Swinh (50 nM) decreased and Jasp (100 nM) increased NO production by PAEC. The stimulatory effect of Jasp on NO production was dependent on the availability of extracellular l-arginine. Our results indicate that the state of actin microfilaments in PAEC regulates l-arginine transport and that this regulation can affect NO production by PAEC.

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