Chloramphenicol-resistant variants of Pseudomonas aeruginosa defective in amino acid transport

1980 ◽  
Vol 58 (10) ◽  
pp. 1165-1171 ◽  
Author(s):  
Jean E. Irvin ◽  
Jordan M. Ingram
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Amino Acid ◽  
Respiratory Activity ◽  
Amino Acid Uptake ◽  
Acid Uptake ◽  
Acid Transport ◽  
Transport Capacity ◽  
Wild Type ◽  
High Concentrations ◽  
High Level

High-level chloramphenicol (CM) resistant variants of Pseudomonas aeruginosa were isolated after culture of the wild-type (WT) strain in broth containing high concentrations of the drug. These variants exhibit reduced ability to accumulate several amino acids. The extent of reduction in transport capacity is a function of the concentration of CM in which the variants are grown. Respiratory activity is not reduced in these strains. Amino acid uptake is not affected by the presence of CM during assay. An isogenic strain carrying a plasmid coding for CM resistance does not show this response to CM. Transport capacity is restored to the WT level in CM-sensitive revertants. These results suggest that the acquisition of CM resistance in P. aeruginosa is associated with a fundamental alteration in membrane permeability which is regulated by metabolism in the presence of the drug. The ramifications of this for the study of CM action and resistance are discussed.

scholarly journals Effect of high altitude on human placental amino acid transport

2020 ◽  
Vol 128 (1) ◽  
pp. 127-133 ◽  
Author(s):  
Owen. R. Vaughan ◽  
Fredrick Thompson ◽  
Ramón. A. Lorca ◽  
Colleen G. Julian ◽  
Theresa L. Powell ◽  
...  
Keyword(s):  
Birth Weight ◽  
Amino Acid ◽  
High Altitude ◽  
Sea Level ◽  
Amino Acid Transport ◽  
Acid Uptake ◽  
Acid Transport ◽  
Transport Capacity ◽  
System A ◽  
Low Altitude

Women residing at high altitudes deliver infants of lower birth weight than at sea level. Birth weight correlates with placental system A-mediated amino acid transport capacity, and severe environmental hypoxia reduces system A activity in isolated trophoblast and the mouse placenta. However, the effect of high altitude on human placental amino acid transport remains unknown. We hypothesized that microvillous membrane (MVM) system A and system L amino acid transporter activity is lower in placentas of women living at high altitude compared with low-altitude controls. Placentas were collected at term from healthy pregnant women residing at high altitude (HA; >2,500 m; n = 14) or low altitude (LA; <1,700 m; n = 14) following planned, unlabored cesarean section. Birth weight, but not placenta weight, was 13% lower in HA pregnancies (2.88 ± 0.11 kg) compared with LA (3.30 ± 0.07 kg, P < 0.01). MVM erythropoietin receptor abundance, determined by immunoblot, was greater in HA than in LA placentas, consistent with lower placental oxygen levels at HA. However, there was no effect of altitude on MVM system A or L activity, determined by Na+-dependent [14C]methylaminoisobutyric acid uptake and [3H]leucine uptake, respectively. MVM abundance of glucose transporters (GLUTs) 1 and 4 and basal membrane GLUT4 were also similar in LA and HA placentas. Low birth weights in the neonates of women residing at high altitude are not a consequence of reduced placental amino acid transport capacity. These observations are in general agreement with studies of IUGR babies at low altitude, in which MVM system A activity is downregulated only in growth-restricted babies with significant compromise. NEW & NOTEWORTHY Babies born at high altitude are smaller than at sea level. Birth weight is dependent on growth in utero and, in turn, placental nutrient transport. We determined amino acid transport capacity in placentas collected from women resident at low and high altitude. Altitude did not affect system A amino acid transport across the syncytiotrophoblast microvillous membrane, suggesting that impaired placental amino acid transport does not contribute to reduced birth weight in this high-altitude population.

scholarly journals Evidence against the participation of the γ-glutamyltransferase-γ-glutamylcylclotransferase pathway in amino acid transport by rabbit erythrocytes

1975 ◽  
Vol 152 (3) ◽  
pp. 713-715 ◽  
Author(s):  
J D Young ◽  
J C Ellory ◽  
P C Wright
Keyword(s):  
Amino Acid ◽  
Amino Acid Transport ◽  
Amino Acid Uptake ◽  
Acid Uptake ◽  
Acid Transport

The GSH concentration of rabbit erythrocytes was monitored under conditions of large net transport of alanine, phenylalane and lysine in the absence of glucose. In no case was there an appreciable alteration in GSH concentration during amino acid uptake. It is suggested that the γ-glutamyltransferase-γ-glutamylcyclotransferase pathway does not participate in amino acid transport by these cells.

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.

Active Transport of α-Aminoisobutyric Acid by the Isolated Midgut of Hyalophora Cecropia

1972 ◽  
Vol 56 (1) ◽  
pp. 167-172
Author(s):  
SIGNE NEDERGAARD
Keyword(s):  
Amino Acid ◽  
Active Transport ◽  
Opposite Direction ◽  
Amino Acid Transport ◽  
Amino Acid Uptake ◽  
Acid Uptake ◽  
Acid Transport ◽  
Active Process ◽  
Aminoisobutyric Acid ◽  
Amino Acid Flux

1. The α-aminoisobutyric acid flux from lumen to blood of the isolated Cecropia midgut is around 17 µmole/h, while the amino acid flux in the opposite direction is on average 0.3 µmole/h. 2. The amino acid uptake is inhibited by lack of oxygen. It is suggested that the amino acid transport from lumen to blood is an active process. 3. The amino acid uptake is inhibited by short-circuiting the midgut potential, indicating that there is no direct correlation between the active transport of potassium and the uptake of the amino acid by the midgut.

Acute acidosis inhibits liver amino acid transport: no primary role for the urea cycle in acid-base balance

1994 ◽  
Vol 267 (6) ◽  
pp. F1015-F1020 ◽  
Author(s):  
L. Boon ◽  
P. J. Blommaart ◽  
A. J. Meijer ◽  
W. H. Lamers ◽  
A. C. Schoolwerth
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Hepatic Vein ◽  
Amino Acid Transport ◽  
Amino Acid Uptake ◽  
Acid Uptake ◽  
Primary Role ◽  
Urea Synthesis ◽  
Acid Transport ◽  
Acid Base

To examine further the role of the liver in acid-base homeostasis, we studied hepatic amino acid uptake and urea synthesis in rats in vivo during acute acidosis and alkalosis, induced by infusion of 1.8 mmol of HCl or NaHCO3 over 3 h. Amino acids and NH4+ were measured in portal vein, hepatic vein, and aortic plasma, and arteriovenous differences of amino acids and urinary urea and NH4+ excretion were measured. In acidosis, urinary urea excretion was reduced 36% (P < 0.01), whereas urinary NH4+ excretion increased ninefold (P < 0.01), but the sum of urea and NH4+ excretion was unchanged. Total hepatic amino acid uptake, as determined from arteriovenous differences, was decreased by 63% (P < 0.01) in acidosis, with the major effect being noted with alanine and glycine. Only glutamine was released in both acidosis and alkalosis but was not significantly different in the two conditions. Since intracellular concentrations of readily transportable amino acids were not different at low pH despite accelerated protein degradation, these results indicate that hepatic amino acid transport was inhibited markedly and sufficiently to explain the observed decrease in urea synthesis. Total hepatic vein amino acid content was greater in acidosis than alkalosis (P < 0.01). Directly or indirectly, by conversion to glutamine elsewhere, these increased amino acids were degraded in kidney and accounted for the ninefold increase in urinary NH4+ excretion.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Use of [11C]Aminocyclohexanecarboxylate for the Measurement of Amino Acid Uptake and Distribution Volume in Human Brain

1990 ◽  
Vol 10 (5) ◽  
pp. 727-739 ◽  
Author(s):  
Robert A. Koeppe ◽  
Thomas Mangner ◽  
A. Lorris Betz ◽  
Barry L. Shulkin ◽  
Richard Allen ◽  
...  
Keyword(s):  
Amino Acid ◽  
Human Brain ◽  
Amino Acid Transport ◽  
Normal Volunteer ◽  
Amino Acid Uptake ◽  
Distribution Volume ◽  
Transport Rate ◽  
Acid Uptake ◽  
Acid Transport ◽  
Influx Rate

A quantitative positron emission tomographic (PET) method to measure amino acid blood–brain barrier (BBB) transport rate and tissue distribution volume (DV) has been developed using 11C-labeled aminocyclohexanecarboxylate (ACHC), a nonmetabolized amino acid analogue. Dynamic PET data were acquired as a series of 15 scans covering a total of 60 min and analyzed by means of a two-compartment, two-parameter model. Functional images were calculated for the amino acid transport rate constants across the BBB and the amino acid DV in the brain. Results show [11C]ACHC to have an influx rate constant in gray matter of ∼0.03–0.04 ml g−1 min−1, indicating a single-pass extraction fraction of ∼5–7%. The intersubject coefficient of variation was ∼15% while intrasubject variability of repeat scans was only slightly greater than 5%. Studies were performed in 15 young normal volunteer control subjects, 5 elderly controls, 7 patients with probable Alzheimer's disease, and one patient with phenylketonuria. Results indicate that [11C]-ACHC will serve as the basis of a method for measuring amino acid transport rate and DV in the normal and pathological human brain.

Amino Acid Uptake by Amino Acid Analog Resistant Tobacco Cell Lines

1978 ◽  
Vol 33 (9-10) ◽  
pp. 634-640 ◽  
Author(s):  
Jochen Berlin ◽  
Jade M. Widholm
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Cell Line ◽  
Transport System ◽  
Amino Acid Uptake ◽  
Resistant Cell ◽  
Resistant Cell Line ◽  
Acid Uptake ◽  
Wild Type ◽  
Phenylalanine Transport

Abstract Two tobacco cell lines resistant to p-fiuorophenylalanine (PFP) and one resistant to 5-methyltryptophan (5-MT) are compared with wild type cells in their ability to absorb amino acids from the medium. One p-fluorophenylalanine-resistant cell line shows greatly reduced uptake of all amino acids so is resistant to growth inhibition by other amino acid analogs. The impaired absorption is noted with amino acids, amino acid analogs and shikimate, but not with cinnamate, salicylate, nicotine, glucose, 3-O-methylglucose and palmitate. The phenylalanine transport system of the PFP-resistant cell line and the wild type both have Km values of 90 µᴍ, but have different V max values. Several analogs of phenylalanine and several neutral L-amino acids inhibit the phenylalanine transport system, while ʟ-aspartic acid, ʟ-arginine, ᴅ-phenylalanine or chlorogenic acid do not interfere with the ʟ-phenylalanine uptake. The results indicate the presence of more than one transport system for amino acid uptake. The lessened uptake of all amino acids, the specificity of the uptake systems and the unchanged binding let us conclude that a pleiotropic mutation or that some inhibitor causes the reduced uptake of all amino acids by the PFP-resistant cell line.

Amino acid uptake profiling of wild type and recombinant Streptomyces lividans TK24 batch fermentations

2011 ◽  
Vol 152 (4) ◽  
pp. 132-143 ◽  
Author(s):  
Pieter-Jan D’Huys ◽  
Ivan Lule ◽  
Sven Van Hove ◽  
Dominique Vercammen ◽  
Christine Wouters ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Uptake ◽  
Streptomyces Lividans ◽  
Acid Uptake ◽  
Wild Type

scholarly journals Quaternary structure of the small amino acid transporter OprG from Pseudomonas aeruginosa

2018 ◽  
Vol 293 (44) ◽  
pp. 17267-17277 ◽  
Author(s):  
Raghavendar Reddy Sanganna Gari ◽  
Patrick Seelheim ◽  
Brendan Marsh ◽  
Volker Kiessling ◽  
Carl E. Creutz ◽  
...  
Keyword(s):  
Amino Acids ◽  
Pseudomonas Aeruginosa ◽  
Amino Acid ◽  
Outer Membrane ◽  
Amino Acid Transport ◽  
Acid Uptake ◽  
Substrate Uptake ◽  
Acid Transport ◽  
Oligomeric State ◽  
Loop Region

Pseudomonas aeruginosa is an opportunistic human pathogen that causes nosocomial infections. The P. aeruginosa outer membrane contains specific porins that enable substrate uptake, with the outer membrane protein OprG facilitating transport of small, uncharged amino acids. However, the pore size of an eight-stranded β-barrel monomer of OprG is too narrow to accommodate even the smallest transported amino acid, glycine, raising the question of how OprG facilitates amino acid uptake. Pro-92 of OprG is critically important for amino acid transport, with a P92A substitution inhibiting transport and the NMR structure of this variant revealing that this substitution produces structural changes in the barrel rim and restricts loop motions. OprG may assemble into oligomers in the outer membrane (OM) whose subunit interfaces could form a transport channel. Here, we explored the contributions of the oligomeric state and the extracellular loops to OprG's function. Using chemical cross-linking to determine the oligomeric structures of both WT and P92A OprG in native outer membranes and atomic force microscopy, and single-molecule fluorescence of the purified proteins reconstituted into lipid bilayers, we found that both protein variants form oligomers, supporting the notion that subunit interfaces in the oligomer could provide a pathway for amino acid transport. Furthermore, performing transport assays with loop-deleted OprG variants, we found that these variants also can transport small amino acids, indicating that the loops are not solely responsible for substrate transport. We propose that OprG functions as an oligomer and that conformational changes in the barrel–loop region might be crucial for its activity.

Sign in / Sign up

Export Citation Format

Share Document