Oxytocin analogues with inhibitory properties, containing in position 2 a hydrophobic amino acid of D-configuration

1985 ◽  
Vol 50 (1) ◽  
pp. 132-145 ◽  
Author(s):  
Michal Lebl ◽  
Tomislav Barth ◽  
Linda Servítová ◽  
Jiřina Slaninová ◽  
Karel Jošt
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Inhibitory Activity ◽  
Structural Features ◽  
Inhibitory Effect ◽  
Hydrophobic Amino Acid

Ten analogues derived from oxytocin, deamino-oxytocin and deamino-carba-oxytocin were prepared which contained a D-amino acid in the position 2 of the parent system. The following D-amino acids were introduced: tyrosine, phenylalanine, p-methylphenylalanine, p-ethylphenylalanine and O-ethyltyrosine. Combination of two structural features which alone lead to strong inhibitors (a suitable D-amino acid in position 2 and a penicillamine moiety in position 1) did not enhance the inhibitory effect. Compounds containing D-tyrosine are weak agonists in the uterotonic assay; in case of 1-carba-analogues they can be converted into sulfoxides with low inhibitory activity. Analogues with D-phenylalanine substituted in the para-position are the most potent antagonists of the uterotonic effect of oxytocin (pA2 = 8.73) in vitro.

EFFECTS OF DIHYDROSTREPTOMYCIN ON AMINO ACID INCORPORATION INTO THE PROTEINS OF M. TUBERCULOSIS (BCG)

1959 ◽  
Vol 37 (1) ◽  
pp. 687-697
Author(s):  
E. Stachiewicz ◽  
J. H. Quastel
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Sodium Stearate ◽  
Ehrlich Ascites Carcinoma ◽  
Inhibitory Effect ◽  
Total Radioactivity ◽  
Amino Acid Incorporation ◽  
Acid Incorporation ◽  
Drug Concentrations

A study has been made of the effects of dihydrostreptomycin on amino acid incorporation into the proteins of M. tuberculosis (BCG). Suspensions of this organism on incubation at 37° with glycine-1-C14give rise, aerobically, to labelled proteins in which 80% of the radioactivity appears in the glycine and serine moieties of the proteins and about 20% in alanine and aspartic acid. In presence of glycine-2-C14, radioactivity appears in a larger number of amino acids of the protein. Incubation with serine-3-C14leads to a distribution of radioactivity in the amino acids in BCG proteins but alanine-1-C14and valine-1-C14give rise to proteins with the radioactivity almost entirely in the corresponding amino acids. The process of aerobic incorporation of radioactivity from glycine-1-C14in BCG proteins is stimulated by the presence of glucose, glycerol, sodium pyruvate, sodium stearate, or sodium benzoate in the medium in which the cells are incubated, the rate of incorporation being approximately constant over a period of 4 hours. The incorporation depends largely on the presence of oxygen. Dihydrostreptomycin (33 μg per ml) markedly inhibits labelling of proteins in the cell suspensions in presence of radioactive amino acids, the inhibition increasing with concentration of the streptomycin to an optimal concentration of 200 μg/ml. Penicillin and isonicotinic hydrazide are inactive but chloromycetin is an effective inhibitor. Cyanide, arsenite, and azide are inhibitory. The presence of lecithin stimulates incorporation of radioactivity from glycine-1-C14into BCG proteins. Dihydrostreptomycin inhibitions of amino acid incorporation into BCG proteins increase with time of incubation of the cells with the drug. Concentrations of dihydrostreptomycin that inhibit labelled amino acid incorporation into labelled proteins by 50% have no effect on BCG respiration. The drug has no inhibitory effect on labelled amino acid incorporation in E. coli or Ehrlich ascites carcinoma cells in vitro but is effective with M. phlei. It does not affect selectively the distribution of radioactivities of the component amino acids of BCG proteins; only the total radioactivity incorporated into the proteins is diminished. The results lead to the conclusion that dihydrostreptomycin brings about an inhibition of protein synthesis in the BCG strain of M. tuberculosis at concentrations at which it exerts antibiotic effects.

Amino acids in the rat intestinal lumen regulate their own absorption from a distant intestinal site

2009 ◽  
Vol 297 (2) ◽  
pp. G292-G298 ◽  
Author(s):  
Fadi H. Mourad ◽  
Kassem A. Barada ◽  
Carmen Khoury ◽  
Tamim Hamdi ◽  
Nayef E. Saadé ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Inhibitory Effect ◽  
Intestinal Lumen ◽  
Jejunal Segment ◽  
Subdiaphragmatic Vagotomy ◽  
Regulatory Loop ◽  
Dependent Transport

Intestinal nutrient transport is altered in response to changes in dietary conditions and luminal substrate level. It is not clear, however, whether an amino acid in the intestinal lumen can acutely affect its own absorption from a distant site. Our aim is to study the effect of an amino acid present in rat small intestinal segment on its own absorption from a proximal or distal site and elucidate the underlying mechanisms. The effect of instillation of alanine (Ala) in either jejunum or ileum on its own absorption at ileal or jejunal level was examined in vivo. The modulation of this intestinal regulatory loop by the following interventions was studied: tetrodotoxin (TTX) added to Ala, subdiaphragmatic vagotomy, chemical ablation of capsaicin-sensitive primary afferent (CSPA) fibers, and IV administration of calcitonin gene-related peptide (CGRP) antagonist. In addition, the kinetics of jejunal Ala absorption and the importance of Na+-dependent transport were studied in vitro after instilling Ala in the ileum. Basal jejunal Ala absorption [0.198 ± 0.018 μmol·cm−1·20 min−1 (means ± SD)] was significantly decreased with the instillation of 20 mM Ala in the ileum or in an adjacent distal jejunal segment (0.12 ± 0.015; P < 0.0001 and 0.138 ± 0.014; P < 0.002, respectively). Comparable inhibition was observed in the presence of proline in the ileum. Moreover, basal Ala absorption from the ileum (0.169 ± 0.025) was significantly decreased by the presence of 20 mM Ala in the jejunum (0.103 ± 0.027; P < 0.01). The inhibitory effect on jejunal Ala absorption was abolished by TTX, subdiaphragmatic vagotomy, neonatal capsaicin treatment, and CGRP antagonism. In vitro studies showed that Ala in the ileum affects Na+-mediated transport and increases Km without affecting Vmax. Intraluminal amino acids control their own absorption from a distant part of the intestine, by affecting the affinity of the Na+-mediated Ala transporter, through a neuronal mechanism that involves CSPA and CGRP.

EFFECTS OF DIHYDROSTREPTOMYCIN ON AMINO ACID INCORPORATION INTO THE PROTEINS OF M. TUBERCULOSIS (BCG)

1959 ◽  
Vol 37 (5) ◽  
pp. 687-697 ◽  
Author(s):  
E. Stachiewicz ◽  
J. H. Quastel
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Sodium Stearate ◽  
Ehrlich Ascites Carcinoma ◽  
Inhibitory Effect ◽  
Total Radioactivity ◽  
Amino Acid Incorporation ◽  
Acid Incorporation ◽  
Drug Concentrations

A study has been made of the effects of dihydrostreptomycin on amino acid incorporation into the proteins of M. tuberculosis (BCG). Suspensions of this organism on incubation at 37° with glycine-1-C14give rise, aerobically, to labelled proteins in which 80% of the radioactivity appears in the glycine and serine moieties of the proteins and about 20% in alanine and aspartic acid. In presence of glycine-2-C14, radioactivity appears in a larger number of amino acids of the protein. Incubation with serine-3-C14leads to a distribution of radioactivity in the amino acids in BCG proteins but alanine-1-C14and valine-1-C14give rise to proteins with the radioactivity almost entirely in the corresponding amino acids. The process of aerobic incorporation of radioactivity from glycine-1-C14in BCG proteins is stimulated by the presence of glucose, glycerol, sodium pyruvate, sodium stearate, or sodium benzoate in the medium in which the cells are incubated, the rate of incorporation being approximately constant over a period of 4 hours. The incorporation depends largely on the presence of oxygen. Dihydrostreptomycin (33 μg per ml) markedly inhibits labelling of proteins in the cell suspensions in presence of radioactive amino acids, the inhibition increasing with concentration of the streptomycin to an optimal concentration of 200 μg/ml. Penicillin and isonicotinic hydrazide are inactive but chloromycetin is an effective inhibitor. Cyanide, arsenite, and azide are inhibitory. The presence of lecithin stimulates incorporation of radioactivity from glycine-1-C14into BCG proteins. Dihydrostreptomycin inhibitions of amino acid incorporation into BCG proteins increase with time of incubation of the cells with the drug. Concentrations of dihydrostreptomycin that inhibit labelled amino acid incorporation into labelled proteins by 50% have no effect on BCG respiration. The drug has no inhibitory effect on labelled amino acid incorporation in E. coli or Ehrlich ascites carcinoma cells in vitro but is effective with M. phlei. It does not affect selectively the distribution of radioactivities of the component amino acids of BCG proteins; only the total radioactivity incorporated into the proteins is diminished. The results lead to the conclusion that dihydrostreptomycin brings about an inhibition of protein synthesis in the BCG strain of M. tuberculosis at concentrations at which it exerts antibiotic effects.

scholarly journals The rfaE Gene from Escherichia coli Encodes a Bifunctional Protein Involved in Biosynthesis of the Lipopolysaccharide Core Precursor ADP-l-glycero-d-manno-Heptose

2000 ◽  
Vol 182 (2) ◽  
pp. 488-497 ◽  
Author(s):  
Miguel A. Valvano ◽  
Cristina L. Marolda ◽  
Mauricio Bittner ◽  
Mike Glaskin-Clay ◽  
Tania L. Simon ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Amino Acid ◽  
Inner Core ◽  
Structural Features ◽  
Predicted Amino Acid Sequence ◽  
E Coli ◽  
K 12 ◽  
Domain I

ABSTRACT The intermediate steps in the biosynthesis of the ADP-l-glycero-d-manno-heptose precursor of inner core lipopolysaccharide (LPS) are not yet elucidated. We isolated a mini-Tn10 insertion that confers a heptoseless LPS phenotype in the chromosome of Escherichia coli K-12. The mutation was in a gene homologous to the previously reported rfaE gene from Haemophilus influenzae. The E. coli rfaE gene was cloned into an expression vector, and an in vitro transcription-translation experiment revealed a polypeptide of approximately 55 kDa in mass. Comparisons of the predicted amino acid sequence with other proteins in the database showed the presence of two clearly separate domains. Domain I (amino acids 1 to 318) shared structural features with members of the ribokinase family, while Domain II (amino acids 344 to 477) had conserved features of the cytidylyltransferase superfamily that includes the aut gene product of Ralstonia eutrophus. Each domain was expressed individually, demonstrating that only Domain I could complement therfaE::Tn10 mutation in E. coli, as well as the rfaE543 mutation ofSalmonella enterica SL1102. DNA sequencing of therfaE543 gene revealed that Domain I had one amino acid substitution and a 12-bp in-frame deletion resulting in the loss of four amino acids, while Domain II remained intact. We also demonstrated that the aut::Tn5 mutation inR. eutrophus is associated with heptoseless LPS, and this phenotype was restored following the introduction of a plasmid expressing the E. coli Domain II. Thus, both domains ofrfaE are functionally different and genetically separable confirming that the encoded protein is bifunctional. We propose that Domain I is involved in the synthesis ofd-glycero-d-manno-heptose 1-phosphate, whereas Domain II catalyzes the ADP transfer to form ADP-d-glycero-d-manno-heptose.

scholarly journals FRUCTOSE-AMINO ACIDS IN LIVER: STIMULI OF AMINO ACID INCORPORATION IN VITRO

1955 ◽  
Vol 215 (1) ◽  
pp. 111-124 ◽  
Author(s):  
Henry Borsook ◽  
Adolph Abrams ◽  
Peter H. Lowy
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Incorporation ◽  
Acid Incorporation

scholarly journals Transdermal Delivery Systems for Ibuprofen and Ibuprofen Modified with Amino Acids Alkyl Esters Based on Bacterial Cellulose

2021 ◽  
Vol 22 (12) ◽  
pp. 6252
Author(s):  
Paula Ossowicz-Rupniewska ◽  
Rafał Rakoczy ◽  
Anna Nowak ◽  
Maciej Konopacki ◽  
Joanna Klebeko ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Bacterial Cellulose ◽  
Acid Ester ◽  
Active Pharmaceutical Ingredient ◽  
Amino Acid Ester ◽  
Isopropyl Ester ◽  
Isopropyl Esters ◽  
Transdermal Application

The potential of bacterial cellulose as a carrier for the transport of ibuprofen (a typical example of non-steroidal anti-inflammatory drugs) through the skin was investigated. Ibuprofen and its amino acid ester salts-loaded BC membranes were prepared through a simple methodology and characterized in terms of structure and morphology. Two salts of amino acid isopropyl esters were used in the research, namely L-valine isopropyl ester ibuprofenate ([ValOiPr][IBU]) and L-leucine isopropyl ester ibuprofenate ([LeuOiPr][IBU]). [LeuOiPr][IBU] is a new compound; therefore, it has been fully characterized and its identity confirmed. For all membranes obtained the surface morphology, tensile mechanical properties, active compound dissolution assays, and permeation and skin accumulation studies of API (active pharmaceutical ingredient) were determined. The obtained membranes were very homogeneous. In vitro diffusion studies with Franz cells were conducted using pig epidermal membranes, and showed that the incorporation of ibuprofen in BC membranes provided lower permeation rates to those obtained with amino acids ester salts of ibuprofen. This release profile together with the ease of application and the simple preparation and assembly of the drug-loaded membranes indicates the enormous potentialities of using BC membranes for transdermal application of ibuprofen in the form of amino acid ester salts.

scholarly journals Phylogenetic analysis and predicted functional effect of protein mutations of E6 and E7 HPV16 strains isolated in Indonesia

2015 ◽  
Vol 24 (4) ◽  
pp. 197-205
Author(s):  
Dwi Wulandari ◽  
Lisnawati Rachmadi ◽  
Tjahjani M. Sudiro
Keyword(s):  
Amino Acids ◽  
Phylogenetic Analysis ◽  
Amino Acid ◽  
Asian American ◽  
Protein Function ◽  
Single Amino Acid ◽  
Hpv16 E6 ◽  
E6 And E7 ◽  
Neutral Mutations

Background: E6 and E7 are oncoproteins of HPV16. Natural amino acid variation in HPV16 E6 can alter its carcinogenic potential. The aim of this study was to analyze phylogenetically E6 and E7 genes and proteins of HPV16 from Indonesia and predict the effects of single amino acid substitution on protein function. This analysis could be used to reduce time, effort, and research cost as initial screening in selection of protein or isolates to be tested in vitro or in vivo.Methods: In this study, E6 and E7 gene sequences were obtained from 12 samples of  Indonesian isolates, which  were compared with HPV16R (prototype) and 6 standard isolates in the category of European (E), Asian (As), Asian-American (AA), African-1 (Af-1), African-2 (Af-2), and North American (NA) branch from Genbank. Bioedit v.7.0.0 was used to analyze the composition and substitution of single amino acids. Phylogenetic analysis of E6 and E7 genes and proteins was performed using Clustal X (1.81) and NJPLOT softwares. Effects of single amino acid substitutions on protein function of E6 and E7 were analysed by SNAP.Results: Java variants and isolate ui66* belonged to European branch, while the others belonged to Asian and African branches. Twelve changes of amino acids were found in E6 and one in E7 proteins. SNAP analysis showed two non neutral mutations, i.e. R10I and C63G in E6 proteins. R10I mutations were found in Af-2 genotype (AF472509) and Indonesian isolates (Af2*), while C63G mutation was found only in Af2*.Conclusion: E6 proteins of HPV16 variants were more variable than E7. SNAP analysis showed that only E6 protein of African-2 branch had functional differences compared to HPV16R.

scholarly journals Identification in skeletal muscle of a distinct extracellular pool of amino acids, and its role in protein synthesis

1971 ◽  
Vol 121 (5) ◽  
pp. 817-827 ◽  
Author(s):  
R. C. Hider ◽  
E. B. Fern ◽  
D. R. London
Keyword(s):  
Skeletal Muscle ◽  
Amino Acids ◽  
Protein Synthesis ◽  
Amino Acid ◽  
Incubation Medium ◽  
Extensor Digitorum Longus ◽  
Extensor Digitorum ◽  
Longus Muscle ◽  
Kinetics Of

1. The kinetics of radioactive labelling of extra- and intra-cellular amino acid pools and protein of the extensor digitorum longus muscle were studied after incubations with radioactive amino acids in vitro. 2. The results indicated that an extracellular pool could be defined, the contents of which were different from those of the incubation medium. 3. It was concluded that amino acids from the extracellular pool, as defined in this study, were incorporated directly into protein.

Amino Acid Transport in a Water-mould: The Possible Regulatory Roles of Calcium and N6-(Δ2-isopentenyl)adenine

1975 ◽  
Vol 53 (9) ◽  
pp. 975-988 ◽  
Author(s):  
Danny P. Singh ◽  
Hérb. B. LéJohn
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Transport ◽  
Osmotic Shock ◽  
Inhibitory Effect ◽  
Transport Systems ◽  
Acid Transport ◽  
Isopentenyl Adenine ◽  
Energy Dependent ◽  
Water Mould

Transport of amino acids in the water-mould Achlya is an energy-dependent process. Based on competition kinetics and studies involving the influence of pH and temperature on the initial transport rates, it was concluded that the 20 amino acids (L-isomers) commonly found in proteins were transported by more than one, possibly nine, uptake systems. This is similar to the pattern elucidated for some bacteria but unlike those uncovered for all fungi studied to date. The nine different transport systems elucidated are: (i) methionine, (ii) cysteine, (iii) proline, (iv) serine–threonine, (v) aspartic and glutamic acids, (vi) glutamine and asparagine, (vii) glycine and alanine, (viii) histidine, lysine, and arginine, and (ix) phenylalanine–tyrosine–tryptophan and leucine–isoleucine–valine as two overlapping groups. Transport of all of these amino acids was inhibited by azide, cyanide, and its derivatives and 2,4-dinitrophenol. These agents normally interfere with metabolism at the level of the electron transport chain and oxidative phosphorylation. Osmotic shock treatment of the cells released, into the shock fluid, a glycopeptide that binds calcium as well as tryptophan but no other amino acid. The shocked cells are incapable of concentrating amino acids, but remain viable and reacquire this capacity when the glycopeptide is resynthesized.Calcium played more than a secondary role in the transport of the amino acids. When bound to the membrane-localized glycopeptide, it permits concentrative transport to take place. However, excess calcium can inhibit transport which can be overcome by chelating with citrate. Calculations show that the concentration of free citrate is most important. At low citrate concentrations (less than 1 mM) in the absence of exogenously supplied calcium, enhancement of amino acid transport occurs. At high concentrations (greater than 5 mM), citrate inhibits but this effect can be reversed by titrating with calcium. Evidently, the glycopeptide acts as a calcium sink to regulate the concentration of calcium made available to the cell for its membrane activities.N6-(Δ2-isopentenyl) adenine (a plant growth 'hormone') and analogues mimic the inhibitory effect of citrate and bind to the glycopeptide as well. Replot data for citrate and N6-(Δ2-isopentyl) adenine inhibition indicate that both agents have no more than one binding constant. These results implicate calcium, glycopeptide, and energy-dependent transport of solutes in some, as yet undefinable, way.

Effect of Cycloheximide on In Vitro and In Vivo Protein Synthesis by Certain Tissues of Rats and Pigeons with Special Reference to Muscle

1973 ◽  
Vol 51 (12) ◽  
pp. 933-941 ◽  
Author(s):  
Njanoor Narayanan ◽  
Jacob Eapen
Keyword(s):  
Amino Acids ◽  
Body Weight ◽  
Protein Synthesis ◽  
Amino Acid ◽  
Amino Acid Incorporation ◽  
Contrast Administration ◽  
Acid Incorporation ◽  
Tissue Homogenates

The effect of cycloheximide in vitro and in vivo on the incorporation of labelled amino acids into protein by muscles, liver, kidneys, and brain of rats and pigeons was studied. In vitro incorporation of amino acids into protein by muscle microsomes, myofibrils, and myofibrillar ribosomes was not affected by cycloheximide. In contrast, administration of the antibiotic into intact animals at a concentration of 1 mg/kg body weight resulted in considerable inhibition of amino acid incorporation into protein by muscles, liver, kidneys, and brain. This inhibition was observed in all the subcellular fractions of these tissues during a period of 10–40 min after the administration of the precursor. Tissue homogenates derived from in vivo cycloheximide-treated animals did not show significant alteration in in vitro amino acid incorporation with the exception of brain, which showed a small but significant enhancement.

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