Search PubMed⌕ Search

Biomedical subjects

E J Lea

Publications and source records attributed to E J Lea.

33 records · Page 2Linked to original sources
PubMed1985

Characterisation of channels induced in planar bilayer membranes by detergent solubilised Escherichia coli porins.

Purified OmpF, OmpC, NmpC, PhoE and Lc (Protein 2) porins from the Escherichia coli outer membrane were incorporated into planar phospholipid bilayer membranes and the permeability properties of the pores studied. Triton X-100 solubilised porin samples showed large and reproducible increases in membrane conductivity composed of discreet single-channel events. The magnitude of the cation selectivity found for the porins was in the order OmpC greater than OmpF greater than NmpC = Lc; PhoE was anion selective. For the cation selective porins the cation/anion permeability ratios in a variety of solutes ranged from 6 to 35. Further information on the internal structure of the porins was obtained by examination of the single-channel conductance and this was used to interpret macroscopic observations and to estimate single-channel diameters. The same porins solubilised in SDS exhibited slight conductance increase with no observable single-channel activity. Use of on-line microcomputer techniques confirmed the ohmic current vs. voltage behaviour for all the single porin channels examined.

Bacterial Outer Membrane Proteins↗
PubMed1983

A new channel-forming antibiotic from Streptomyces coelicolor A3(2) which requires calcium for its activity.

A recently discovered antibiotic (CDA; calcium-dependent antibiotic) of Streptomyces coelicolor A3(2) was found to be effective against a wide range of Gram-positive bacteria only in the presence of calcium ions. Producer and non-producer strains were identified and several media tested for their ability to support antibiotic production. The action of calcium was not simulated by any of the other cations tested. The antibiotic was found to induce discrete conductance fluctuations in planar lipid bilayer consistent with a channel-forming action. The electrical potential difference caused by a concentration difference of various salts across the CDA-containing bilayer, showed the channel to be cation-selective but of a size that discriminated against tetramethyl ammonium and choline ions. The data indicate that the antibiotic activity of CDA is due to its action as a calcium-dependent ionophore.

Anti-Bacterial Agents↗
PubMed1982

The fatty acids of erythrocytes of myocardial infarction patients.

Arachidonic acid (C20 : 4 omega 6) and eicosapentaenoic acid (C20 : 5 omega 3) are precursors of two different series of prostaglandins important in homeostasis of the cardiovascular system. The levels of these and other fatty acids have been measured in a group of 20 patients who had suffered myocardial infarction (samples taken within 12 h of infarction) and a group of 17 healthy age-matched controls using the erythrocyte as a lipid probe. There was no significant difference between the level of C20 : 4 omega 6 in patients and controls. There was however a highly significant difference in the level of the peak containing C20 : 5 omega 3 i.e. 6.60% +/- 0.29 (mean and SE of mean) for controls; 3.91 +/- 0.36 (mean and SE of mean) for patients with myocardial infarction. In each of the two groups the relationship between the levels of the two fatty acids arachidonic acid, eicosapentaenoic acid and their essential fatty acid precursors has been investigated. No significant functional relationship was found except between C18 : 3 omega 3 and C20 : 5 omega 3 in the control group. These results are discussed in relation to homeostasis and recent evidence that levels of these fatty acids can be altered by dietary manipulation.

Arachidonic Acid↗
PubMed1978

The interaction of detergents with bilayer lipid membranes.

Detergents are widely used for extracting and purifying membrane proteins. Four such detergents have been studied to find the extent to which they alone can alter black lipid film conductances. The slope of the plot of conductivity versus concentration for Triton X-100 is 4.54 in the range 0.025--0.15 mM; dodecyl sulphate 0.82 in the range 0.01--1 mM; sodium deoxycholate 1.03 in the range 0.01--1 mM and sodium cholate 1.37 in the range 0.1--10 mM. These ranges are below the respective critical micelle concentrations; above these concentrations the membranes break. Bilayer lipid membrane conductivity measured at constant detergent concentration increases with the conductivity of the bathing salt solution with a slope greater than 1, indicating an effect on the putative pore structures induced by detergents.

Cholic Acids↗
PubMed1975

The effects of the membrane-penetrating polypeptide segment of the human erythrocyte MN-glycoprotein on the permeability of model lipid membranes.

The insoluble peptide, T(is), prepared by trypsin hydrolysis of the MN-glycoprotein (glycophorin) of the human erythrocyte has been incorporated into phospholipid membranes in the form of liposomes and black lipid membranes. The permeability of liposome membranes to 42K+ and of black lipid membranes to water and ions is increased significantly by the presence of the T(is) peptide. Electrophoresis measurements indicate that these effects are not due to the T(is) peptide carrying a net charge. The results suggest that the peptide causes local disordering of the bilayer membrane structures. This is considered in the light of findings published elsewhere: that the MN-glycoprotein penetrates through the cell membrane via a non-polar segment of its polypeptide chain, which is contained intact within the T(is) peptide; that the T(is) peptide is partially helical when associated with phospholipid and forms multimeric 8.0 nm structures within the hydrophobic plane of phospholipid bilayers.

Biological Transport↗
PubMed1974

Ionic relations and electrical properties of isolated oocytes of Bufo bufo with special reference to removal of extracellular chloride.

1. The efflux rate coefficients of sodium, potassium and chloride from immature oocytes of the toad Bufo bufo have been measured and found to be respectively, 0.0069, 0.0023 and 0.0034 min(-1).2. Replacement of external chloride by sulphate resulted in a fall in chloride efflux of about 50%. This has been interpreted in terms of exchange diffusion.3. The membrane conductance has been measured and found to be 0.14 x 10(-3) mhos cm(-2) in good agreement with the value calculated from the flux data, 0.12 x 10(-3) mhos cm(-2), after taking account of the chloride exchange diffusion fraction.

Animals↗
PubMed1967

Partition of sodium fluxes in isolated toad oocytes.

1. The rate constant for Na efflux from the oocyte calculated from (d/dt) (ln [Na(*)](i)]) is only approximately 52% of that calculated from (d/dt)[(ln(d[Na(*)](i))dt)]. The difference may be interpreted by supposing that 48% of the internal Na of the oocyte is either bound to proteins or sequestered in cell organelles.2. The mean rate constant for Na efflux was 6.4 x 10(-3) min(-1) corresponding to an apparent Na efflux rate of 13.3 p-mole/cm(2).sec. When this is corrected for the increase in surface area produced by microvilli the true efflux rate is 1.1-1.3 p-mole/cm(2).sec.3. The action of ouabain (1-5 muM) appears to involve two different effects: (a) there is 48-65% inhibition of the membrane Na pump, and (b) there is a release of some of the sequestered Na in the cell.4. Removal of external K causes a 40% reduction in Na efflux although this value may be an underestimation owing to the presence of K which has leaked from the cell and may be retained near the cell surface.5. Raising the external K concentration to 15 mM reduces the inhibitory effect of ouabain by approximately a half.6. It was concluded that the Na pump in the toad oocyte may have a slightly lower level of activity than that in frog muscle, but that its general properties are similar to those in frog muscle and some other animal cells.

Animals↗