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921.
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Targeting of porin to the outer membrane of Escherichia coli. Rate of trimer assembly and identification of a dimer intermediate 总被引:14,自引:0,他引:14
J Reid H Fung K Gehring P E Klebba H Nikaido 《The Journal of biological chemistry》1988,263(16):7753-7759
Porin, a transmembrane protein in the outer membrane of Escherichia coli, exists in a trimeric structure which is not dissociated during sodium dodecyl sulfate-polyacrylamide gel electrophoresis at 25 degrees C. This unusual stability was utilized in the study of the conformational changes which accompany the targeting of porin to the outer membrane. A delay of 16-44 s between completion of synthesis of a monomer and its assembly into a trimer was found from the ratio of monomers to trimers found in exponentially growing cells. Pulse-chase experiments showed that rapid processing of precursor OmpF molecules was followed by assembly into sodium dodecyl sulfate-resistant oligomers with a half-time of 20 s at 30 degrees C. An intermediate in assembly was isolated by immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis below 10 degrees C and was identified as a metastable dimer. 相似文献
926.
The cause of the pregnancy condition preeclampsia (PE) is thought to be endothelial dysfunction caused by oxidative stress. As abnormal glucose tolerance has also been associated with PE, we use a fluorinated-mimic of this metabolite to establish whether any oxidative damage to lipids and proteins in the erythrocyte membrane has increased cell membrane permeability. Data were acquired using 19F Dynamic-NMR (DNMR) to measure exchange of 3-fluoro-3-deoxyglucose (3-FDG) across the membrane of erythrocytes from 10 pregnant women (5 healthy control women, and 5 from women suffering from PE). Magnetisation transfer was measured using the 1D selective inversion and 2D EXSY pulse sequences, over a range of time delays. Integrated intensities from these experiments were used in matrix diagonalisation to estimate the values of the rate constants of exchange and membrane permeability. No significant differences were observed for the rate of exchange of 3-FDG and membrane permeability between healthy pregnant women and those suffering from PE, leading us to conclude that no oxidative damage had occurred at this carrier-protein site in the membrane. 相似文献
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928.
Tarun K. Dam Manju Sarkar Jharna Ghosal Amalesh Choudhury 《Molecular and cellular biochemistry》1992,117(1):1-9
Control Analysis has been carried out in the first steps of a rat liver glycolytic system. Attention has been focused on the effect of several glucose concentrations on the control, particularly regarding the role of glucokinase. From kinetic studies of the whole metabolic system we have obtained information on the flux variation under different glucose concentrations. This information together with the kinetics of glucokinase has allowed us to calculate Flux Control and Elasticity Coefficients for glucokinase and the Response Coefficient of the system with respect to glucose. The changes in of the value of Flux Control Coefficients demonstrates that in conditions of low glucose concentration, glucokinase is the main enzyme in controlling the flux through the pathway, but at high glucose concentration the control moves to phosphofructokinase. Next, we have compared our results with those obtained with the shortening and titration method, previously described (Torres, N.V., Mateo, F., Mélendez-Hevia, E. and Kacser, H., (1986) Biochem. J. 234, 169–174; Torres, N.V. and Meléndez-Hevia, E. 1991. Molec. Cell. Biochem. 101, 1–10). Furthermore, from knowledge of the enzyme kinetics of the system we have been able to build a model of the pathway that allows us computer similation of its behavior and calculation of the Flux Control Coefficient profile at different glucose concentrations. By the three methods the results correlate, supporting the use of the pathway substrate as external modulator of the metabolic system as a tool for practical application of Control Analysis. 相似文献
929.
T. R. FRASIER T. RASTOGI M. W. BROWN P. K. HAMILTON S. D. KRAUS B. N. WHITE 《Molecular ecology resources》2006,6(4):1025-1029
A North Atlantic right whale (Eubalaena glacialis) genomic library was developed and screened with a (GATA)8 probe to identify tetranucleotide microsatellite loci. Sixteen characterized loci were polymorphic in North Atlantic and/or South Atlantic (Eubalaena australis) right whales, 12 being polymorphic in E. glacialis, and 15 in E. australis. Fourteen of these were combined with 21 other previously identified loci for a suite of 35 loci which can be used to increase resolution of genetic analyses of these species. Multiplex reactions were developed for genotyping samples at these loci, providing a method that is rapid, reliable and cost‐effective. 相似文献
930.
The microbial segment of food webs plays a crucial role in lacustrine food-web functioning and carbon transfer, thereby influencing carbon storage and CO2 emission and uptake in freshwater environments. Variability in microbial carbon processing (autotrophic and heterotrophic production and respiration based on glucose) with depth was investigated in eutrophic, methane-rich Lake Rotsee, Switzerland. In June 2011, 13C-labelling experiments were carried out at six depth intervals in the water column under ambient light as well as dark conditions to evaluate the relative importance of (chemo)autotrophic, mixotrophic and heterotrophic production. Label incorporation rates of phospholipid-derived fatty acid (PLFA) biomarkers allowed us to differentiate between microbial producers and calculate group-specific production. We conclude that at 6 m, net primary production (NPP) rates were highest, dominated by algal photoautotrophic production. At 10 m —the base of the oxycline— a distinct low-light community was able to fix inorganic carbon, while in the hypolimnion, heterotrophic production prevailed. At 2 m depth, high label incorporation into POC could only be traced to nonspecific PLFA, which prevented definite identification, but suggests cyanobacteria as dominating organisms. There was also depth zonation in extracellular carbon release and heterotrophic bacterial growth on recently fixed carbon. Large differences were observed between concentrations and label incorporation of POC and biomarkers, with large pools of inactive biomass settling in the hypolimnion, suggesting late-/post-bloom conditions. Net primary production (115 mmol C m?2 d?1) reached highest values in the epilimnion and was higher than glucose-based production (3.3 mmol C m?2 d?1, highest rates in the hypolimnion) and respiration (5.9 mmol C m?2 d?1, highest rates in the epilimnion). Hence, eutrophic Lake Rotsee was net autotrophic during our experiments, potentially storing large amounts of carbon. 相似文献