Studies on the interaction of Escherichia coli endotoxin with erythrocyte membranes

1982 ◽  
Vol 60 (7) ◽  
pp. 977-985 ◽  
Author(s):  
David V. Godin ◽  
John M. Tuchek ◽  
Maureen E. Garnett
Keyword(s):  
Escherichia Coli ◽  
Membrane Lipids ◽  
Effect Of Temperature ◽  
Erythrocyte Membranes ◽  
Simple Relationship ◽  
In Vitro Effects ◽  
The Stability ◽  
Erythrocyte Stability

Escherichia coli lipopolysaccharide (endotoxin) alters the stability of erythrocytes to hypotonic lysis although the nature and magnitude of the effect varied with temperature and with the type of red blood cell examined. Evidence has been obtained suggesting a possible modulatory role of membrane lipids in governing the molecular consequences of membrane–endotoxin interaction. The marked effect of temperature on the stabilization of red cells by endotoxin was not attributable to variations in toxin binding and was not observed with more conventional structurally unrelated antihemolytic agents. Although chemical modifications which alter the toxicity of endotoxin in vivo also modify its ability to stabilize erythrocytes in vitro, no simple relationship between in vivo endotoxin toxicity and in vitro effects on erythrocyte stability was apparent. The critical dependence of endotoxin antihemolytic effects in vitro on molecular structure may offer a convenient means of assessing the homogeneity of these preparations before performing experiments in vivo.

Applicability of Instability Index for In vitro Protein Stability Prediction

2019 ◽  
Vol 26 (5) ◽  
pp. 339-347 ◽  
Author(s):  
Dilani G. Gamage ◽  
Ajith Gunaratne ◽  
Gopal R. Periyannan ◽  
Timothy G. Russell
Keyword(s):  
Protein Stability ◽  
Caulobacter Crescentus ◽  
Effect Of Temperature ◽  
Instability Index ◽  
Dipeptide Composition ◽  
Experimental Conditions ◽  
The Stability ◽  
Vitro Protein

Background: The dipeptide composition-based Instability Index (II) is one of the protein primary structure-dependent methods available for in vivo protein stability predictions. As per this method, proteins with II value below 40 are stable proteins. Intracellular protein stability principles guided the original development of the II method. However, the use of the II method for in vitro protein stability predictions raises questions about the validity of applying the II method under experimental conditions that are different from the in vivo setting. Objective: The aim of this study is to experimentally test the validity of the use of II as an in vitro protein stability predictor. Methods: A representative protein CCM (CCM - Caulobacter crescentus metalloprotein) that rapidly degrades under in vitro conditions was used to probe the dipeptide sequence-dependent degradation properties of CCM by generating CCM mutants to represent stable and unstable II values. A comparative degradation analysis was carried out under in vitro conditions using wildtype CCM, CCM mutants and two other candidate proteins: metallo-β-lactamase L1 and α -S1- casein representing stable, borderline stable/unstable, and unstable proteins as per the II predictions. The effect of temperature and a protein stabilizing agent on CCM degradation was also tested. Results: Data support the dipeptide composition-dependent protein stability/instability in wt-CCM and mutants as predicted by the II method under in vitro conditions. However, the II failed to accurately represent the stability of other tested proteins. Data indicate the influence of protein environmental factors on the autoproteolysis of proteins. Conclusion: Broader application of the II method for the prediction of protein stability under in vitro conditions is questionable as the stability of the protein may be dependent not only on the intrinsic nature of the protein but also on the conditions of the protein milieu.

scholarly journals In Vitro and In Vivo Assessment of the Potential of Escherichia coli Phages to Treat Infections and Survive Gastric Conditions

2021 ◽  
Vol 9 (9) ◽  
pp. 1869
Author(s):  
Joanna Kaczorowska ◽  
Eoghan Casey ◽  
Gabriele A. Lugli ◽  
Marco Ventura ◽  
David J. Clarke ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Galleria Mellonella ◽  
Enterotoxigenic Escherichia Coli ◽  
E Coli ◽  
Composite Mixture ◽  
Ph Conditions ◽  
The Stability ◽  
Higher Sensitivity

Enterotoxigenic Escherichia coli (ETEC) and Shigella ssp. infections are associated with high rates of mortality, especially in infants in developing countries. Due to increasing levels of global antibiotic resistance exhibited by many pathogenic organisms, alternative strategies to combat such infections are urgently required. In this study, we evaluated the stability of five coliphages (four Myoviridae and one Siphoviridae phage) over a range of pH conditions and in simulated gastric conditions. The Myoviridae phages were stable across the range of pH 2 to 7, while the Siphoviridae phage, JK16, exhibited higher sensitivity to low pH. A composite mixture of these five phages was tested in vivo in a Galleria mellonella model. The obtained data clearly shows potential in treating E. coli infections prophylactically.

scholarly journals Binding of the Escherichia coli cyclic AMP receptor protein to DNA fragments containing consensus nucleotide sequences

1989 ◽  
Vol 261 (2) ◽  
pp. 649-653 ◽  
Author(s):  
K Gaston ◽  
A Kolb ◽  
S Busby
Keyword(s):  
Escherichia Coli ◽  
Cyclic Amp ◽  
Inverse Correlation ◽  
Nucleotide Sequences ◽  
Receptor Protein ◽  
Dna Fragments ◽  
Naturally Occurring ◽  
The Stability

Binding of the Escherichia coli CRP protein to DNA fragments carrying nucleotide sequences closely corresponding to the consensus is very tight with a dissociation time of over 2 h in our conditions. The concentration of cyclic AMP required for this binding is below the physiological range of intracellular cyclic AMP concentrations. Changes in nucleotide sequence at positions that are not well-conserved between different naturally-occurring CRP sites allow a more rapid dissociation of CRP-DNA complexes. There is an inverse correlation between the stability of CRP binding to sites in vitro and the repression by glucose of expression dependent on these sites in vivo: expression that is dependent on the tighter binding sites cannot be repressed by the inclusion of glucose in the growth medium.

scholarly journals (p)ppGpp Inhibits Polynucleotide Phosphorylase from Streptomyces but Not from Escherichia coli and Increases the Stability of Bulk mRNA in Streptomyces coelicolor

2010 ◽  
Vol 192 (17) ◽  
pp. 4275-4280 ◽  
Author(s):  
Marcha L. Gatewood ◽  
George H. Jones
Keyword(s):  
Escherichia Coli ◽  
Chemical Stability ◽  
Antibiotic Production ◽  
Streptomyces Coelicolor ◽  
Polynucleotide Phosphorylase ◽  
Streptomyces Antibioticus ◽  
E Coli ◽  
The Stability

ABSTRACT ppGpp regulates gene expression in a variety of bacteria and in plants. We proposed previously that ppGpp or its precursor, pppGpp [referred to collectively as (p)ppGpp], or both might regulate the activity of the enzyme polynucleotide phosphorylase in Streptomyces species. We have examined the effects of (p)ppGpp on the polymerization and phosphorolysis activities of PNPase from Streptomyces coelicolor, Streptomyces antibioticus, and Escherichia coli. We have shown that (p)ppGpp inhibits the activities of both Streptomyces PNPases but not the E. coli enzyme. The inhibition kinetics for polymerization using the Streptomyces enzymes are of the mixed noncompetitive type, suggesting that (p)ppGpp binds to a region other than the active site of the enzyme. ppGpp also inhibited the phosphorolysis of a model RNA substrate derived from the rpsO-pnp operon of S. coelicolor. We have shown further that the chemical stability of mRNA increases during the stationary phase in S. coelicolor and that induction of a plasmid-borne copy of relA in a relA-null mutant increases the chemical stability of bulk mRNA as well. We speculate that the observed inhibition in vitro may reflect a role of ppGpp in the regulation of antibiotic production in vivo.

Zum Wirkungsmechanismus von 1-Nitroso-3-nitro-1-methylguanidin (NNMG) bei der Mutationsauslösung / The Mode of Action of 1-Nitroso-3-nitro-1-methyl-guanidine (NNMG) in Mutagenesis

1969 ◽  
Vol 24 (7) ◽  
pp. 903-910 ◽  
Author(s):  
R. Süssmuth ◽  
F. Lingens
Keyword(s):  
Escherichia Coli ◽  
Citric Acid ◽  
Nucleic Acids ◽  
Mutation Rate ◽  
Control Experiment ◽  
Ph Dependence ◽  
The Stability ◽  
Escherichia Coli B

1. The stability of NNMG with respect to pH was investigated. A maximum, with a half-life of 40 hours, was found in phosphate-citric acid buffer at pH 5. The stability decreases quickly with increasing concentration of hydroxyl ions. At the optimal mutation rate (pH 6, 37°, and shaking) the half-live is only 14,4 hours.2. NNMG uptake (using labelled NNMG) and mutation rate in the range pH 3,5 — 8,0 show an optimum at pH 6,0/6,5 for both Escherichia coli B and the red ad- mutant E 188 of Saccharomyces cerevisiae. The pH-dependence of NNMG uptake and mutation rate is similar.3. The methylation of nucleic acids by means of [3H]-methyl-NNMG or [14C] -methyl-NNMG in phosphate-citric acid at 37° while shaking increases with the growing concentration of hydroxyl ions. At the optimal mutation rate, however, NNMG methylates relatively poor.4. Incorporation of radioactive NNMG into Escherichia coli B results in labelling of nucleic acids and proteins. The labelling of DNA shows an optimum at pH 6. The extent of methylation was found to be higher in vivo than in vitro.5. In the presence of cysteine or β-mercaptoethanol nucleic acids are methylated more intensively than in the cysteine-free control experiment. After one hour the methylation was about twentyfold higher in the presence of cysteine or β-mercaptoethanol compared to the cysteine-free control experiment. The methylation in presence of SH-compounds shows a maximum at pH 6. The explanation of both a higher methylation rate and its optimum at pH 6 suggests the activation of methylation by means of sulphhydryl-groups.

scholarly journals Correlation of In Vivo and In Vitro Phase Transitions of Membrane Lipids in Escherichia coli

1970 ◽  
Vol 67 (2) ◽  
pp. 606-612 ◽  
Author(s):  
P. Overath ◽  
H. U. Schairer ◽  
W. Stoffel
Keyword(s):  
Escherichia Coli ◽  
Phase Transitions ◽  
Membrane Lipids

In Vivo and In Vitro Effects of Tea Extracts on Enterotoxigenic Escherichia coli-induced Intestinal Fluid Loss in Animal Models

2006 ◽  
Vol 43 (4) ◽  
pp. 459-469 ◽  
Author(s):  
M.J. Bruins ◽  
R. Cermak ◽  
J.L. Kiers ◽  
J. van der Meulen ◽  
J.M.M. van Amelsvoort ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Animal Models ◽  
Enterotoxigenic Escherichia Coli ◽  
Fluid Loss ◽  
Intestinal Fluid ◽  
In Vitro Effects

scholarly journals Self-assembly of spectrin oligomers in vitro: a basis for a dynamic cytoskeleton.

1981 ◽  
Vol 88 (2) ◽  
pp. 463-468 ◽  
Author(s):  
J S Morrow ◽  
V T Marchesi
Keyword(s):  
Self Assembly ◽  
Erythrocyte Membranes ◽  
Membrane Structure And Function ◽  
High Concentrations ◽  
The Stability ◽  
And Function ◽  
Low Ionic Strength ◽  
Affinity Interaction

Purified human erythrocyte spectrin is able to form large oligomeric species without the collaboration of any other proteins. This reversible self-assembly process is both temperature and concentration dependent and seems to be mediated by the same kinds of low affinity noncovalent associations between spectrin monomers that promote tetramer formation. Low ionic strength extracts of erythrocyte membranes also contain these oligomeric species. These results support the idea that spectrin oligomers and the factors that regulate their formation may be responsible for both the stability and the versatility of the erythrocyte membrane cytoskeleton. It is postulated that the high concentrations of spectrin necessary for oligomerization are maintained in vivo by a high-affinity interaction with ankyrin. Such a coupling of high and low affinity interactions in multifunctional proteins may have significant implications for membrane structure and function.

In Vivo and in Vitro Effects of a Novel Enterotoxin STb Produced by Escherichia coli

1986 ◽  
Vol 153 (5) ◽  
pp. 893-901 ◽  
Author(s):  
C. S. Weikel ◽  
H. N. Nellans ◽  
R. L. Guerrant
Keyword(s):  
Escherichia Coli ◽  
In Vitro Effects
Sign in / Sign up

Export Citation Format

Share Document