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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
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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
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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
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A RAPD marker specific for the G genome of wheat was identified. The corresponding 1171-bp DNA sequence was cloned and analyzed. Screening of the database did not reveal any homologies with the known plant DNA sequences. Using the primers specific to the flanking regions of the marker sequence, PCR analysis of the polyploid wheat species and the diploid species of the section Sitopsis was carried out. In addition, using the cloned sequence as a molecular hybridization probe, RFLP analysis of the genomic DNA of these species was performed. 相似文献
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