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1.
Julie K. De Zutter Kara B. Levine Di Deng Anthony Carruthers 《The Journal of biological chemistry》2013,288(28):20734-20744
The human blood-brain barrier glucose transport protein (GLUT1) forms homodimers and homotetramers in detergent micelles and in cell membranes, where the GLUT1 oligomeric state determines GLUT1 transport behavior. GLUT1 and the neuronal glucose transporter GLUT3 do not form heterocomplexes in human embryonic kidney 293 (HEK293) cells as judged by co-immunoprecipitation assays. Using homology-scanning mutagenesis in which GLUT1 domains are substituted with equivalent GLUT3 domains and vice versa, we show that GLUT1 transmembrane helix 9 (TM9) is necessary for optimal association of GLUT1-GLUT3 chimeras with parental GLUT1 in HEK cells. GLUT1 TMs 2, 5, 8, and 11 also contribute to a less abundant heterocomplex. Cell surface GLUT1 and GLUT3 containing GLUT1 TM9 are 4-fold more catalytically active than GLUT3 and GLUT1 containing GLUT3 TM9. GLUT1 and GLUT3 display allosteric transport behavior. Size exclusion chromatography of detergent solubilized, purified GLUT1 resolves GLUT1/lipid/detergent micelles as 6- and 10-nm Stokes radius particles, which correspond to GLUT1 dimers and tetramers, respectively. Studies with GLUTs expressed in and solubilized from HEK cells show that HEK cell GLUT1 resolves as 6- and 10-nm Stokes radius particles, whereas GLUT3 resolves as a 6-nm particle. Substitution of GLUT3 TM9 with GLUT1 TM9 causes chimeric GLUT3 to resolve as 6- and 10-nm Stokes radius particles. Substitution of GLUT1 TM9 with GLUT3 TM9 causes chimeric GLUT1 to resolve as a mixture of 6- and 4-nm particles. We discuss these findings in the context of determinants of GLUT oligomeric structure and transport function. 相似文献
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Vph6 Mutants of Saccharomyces Cerevisiae Require Calcineurin for Growth and Are Defective in Vacuolar H(+)-Atpase Assembly 总被引:1,自引:0,他引:1 下载免费PDF全文
C. S. Hemenway K. Dolinski M. E. Cardenas M. A. Hiller E. W. Jones J. Heitman 《Genetics》1995,141(3):833-844
We have characterized a Saccharomyces cerevisiae mutant strain that is hypersensitive to cyclosporin A (CsA) and FK506, immunosuppressants that inhibit calcineurin, a serine-threonine-specific phosphatase (PP2B). A single nuclear mutation, designated cev1 for calcineurin essential for viability, is responsible for the CsA-FK506-sensitive phenotype. The peptidyl-prolyl cis-trans isomerases cyclophilin A and FKBP12, respectively, mediate CsA and FK506 toxicity in the cev1 mutant strain. We demonstrate that cev1 is an allele of the VPH6 gene and that vph6 mutant strains fail to assemble the vacuolar H(+)-ATPase (V-ATPase). The VPH6 gene was mapped on chromosome VIII and is predicted to encode a 181-amino acid (21 kD) protein with no identity to other known proteins. We find that calcineurin is essential for viability in many mutant strains with defects in V-ATPase function or vacuolar acidification. In addition, we find that calcineurin modulates extracellular acidification in response to glucose, which we propose occurs via calcineurin regulation of the plasma membrane H(+)-ATPase PMA1. Taken together, our findings suggest calcineurin plays a general role in the regulation of cation transport and homeostasis. 相似文献
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Moutassem Billah Masmoudi Lamya Chaoui Nur Eda Topçu Pachka Hammami Mohamed Hichem Kara Didier Aurelle 《Ecology and evolution》2016,6(24):8665-8678
Understanding the factors explaining the observed patterns of genetic diversity is an important question in evolutionary biology. We provide the first data on the genetic structure of a Mediterranean octocoral, the yellow gorgonian Eunicella cavolini, along with insights into the demographic history of this species. We sampled populations in four areas of the Mediterranean Sea: continental France, Algeria, Turkey, and the Balearic and Corsica islands. Along French coasts, three sites were sampled at two depths (20 and 40 m). We demonstrated a high genetic structure in this species (overall FST = 0.13), and most pairwise differentiation tests were significant. We did not detect any difference between depths at the same site. Clustering analyses revealed four differentiated groups corresponding to the main geographical areas. The levels of allelic richness and heterozygosity were significantly different between regions, with highest diversity in Algeria and lowest levels in Turkey. The highest levels of private allelic richness were observed in Algeria followed by Turkey. Such contrasted patterns of genetic diversity were not observed in other Mediterranean octocorals and could be the result of different evolutionary histories. We also provide new empirical evidence of contrasting results between tests and model‐based studies of demographic history. Our results have important consequences for the management of this species. 相似文献
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Short-term effects of wildfire on microbial biomass and abundance in black pine plantation soils in Turkey 总被引:1,自引:0,他引:1
Measurement of soil microbial biomass and abundance offers a means of assessing the response of all microbial populations to changes in the soil environment after a fire. We examined the effects of wildfire on microbial biomass C and N, and abundance of bacteria and fungi 2 months after a fire in a pine plantation. Soil organic carbon (Corg), total nitrogen (Ntot), and electrical conductivity (EC) increased following the fire. In terms of microbial abundance, the overall results showed that burned forest soils had the most bacteria and fungi. Microbial biomass C and N from soil in the burned forest were not significantly different from their unburned forest counterparts. However, microbial indices indicated that fire affects soil microbial community structure by modifying the environmental conditions. The results also suggested that low-intensity fire promotes microorganism functional activity and improves the chemical characteristics of soils under humid climatic conditions. 相似文献
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K E Wilson Z Li M Kara K L Gardner D D Roberts 《Journal of immunology (Baltimore, Md. : 1950)》1999,163(7):3621-3628
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Özgür Aykut Kara Altan Gökşen Tosun Nazan Tekin Şaban Gökçe İsa 《Molecular biology reports》2021,48(4):3439-3449
Molecular Biology Reports - Heat shock protein 90 (Hsp90) is a key chaperone that is abnormally expressed in cancer cells, and therefore, designing novel compounds to inhibit chaperone activities... 相似文献
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Computational fluid dynamics (CFD) modelling based on a commercial package, FLUENT, has been used in the present study. The primary aim of this study is to develop a novel implant by employing CFD techniques. Firstly, CFD analyses on the best design commercially available, which is the Ahmed Glaucoma Valve (AGV®), are accomplished. In the light of the results, the new design focus is selected as the valve. The new design is analysed using GAMBIT and FLUENT software. CFD analyses of the new design and the AGV® are compared and the strengths of the new design are revealed. The results are also compared with the experimental studies AGV® in the literature. It is deduced that the proposed model shows a nonlinear pressure drop response, which is quite similar to that of AGV®. The optimum combination would be a flow rate of 2.5 μl/min and a pressure drop of 1054.58 Pa for the proposed model. 相似文献