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91.
Lookene A Zhang L Tougu V Olivecrona G 《The Journal of biological chemistry》2003,278(39):37183-37194
Lipoprotein lipase (LPL) is dependent on apolipoprotein CII (apoCII), a component of plasma lipoproteins, for function in vivo. The hydrophobic fluorescent probe 1,1'-bis(anilino)-4,4'-bis(naphthalene)-8,8'-disulfonate (bis-ANS) was found to be a potent inhibitor of LPL. ApoCII prevented the inhibition by bis-ANS, and was also able to restore the activity of inhibited LPL in a competitive manner, but only with triacylglycerols with acyl chains longer than three carbons. Studies of fluorescence and surface plasmon resonance indicated that LPL has an exposed hydrophobic site for binding of bis-ANS. The high affinity interaction was characterized by an equilibrium constant Kd of 0.10-0.26 microm and by a relatively high on rate constant kass = 2.0 x 10(4) m(-1) s(-1) and a slow off-rate with a dissociation rate constant kdiss = 1.2 x 10(-4) s(-1). The high affinity binding of bis-ANS did not influence interaction of LPL with heparin or with lipid/water interfaces and did not dissociate the active LPL dimer into monomers. Analysis of fragments of LPL after photoincorporation of bis-ANS indicated that the high affinity binding site was located in the middle part of the N-terminal folding domain. We propose that bis-ANS binds to an exposed hydrophobic area that is located close to the active site. This area may be the binding site for individual substrate molecules and also for apoCII. 相似文献
92.
Constance Mehlgarten Daniel Jablonowski Uta Wrackmeyer Susan Tschitschmann David Sondermann Gunilla Jäger Zhizhong Gong Anders S. Byström Raffael Schaffrath Karin D. Breunig 《Molecular microbiology》2010,76(5):1082-1094
Based on studies in yeast and mammalian cells the Elongator complex has been implicated in functions as diverse as histone acetylation, polarized protein trafficking and tRNA modification. Here we show that Arabidopsis mutants lacking the Elongator subunit AtELP3/ELO3 have a defect in tRNA wobble uridine modification. Moreover, we demonstrate that yeast elp3 and elp1 mutants expressing the respective Arabidopsis Elongator homologues AtELP3/ELO3 and AtELP1/ELO2 assemble integer Elongator complexes indicating a high degree of structural conservation. Surprisingly, in vivo complementation studies based on Elongator‐dependent tRNA nonsense suppression and zymocin tRNase toxin assays indicated that while AtELP1 rescued defects of a yeast elp1 mutant, the most conserved Elongator gene AtELP3, failed to complement an elp3 mutant. This lack of complementation is due to incompatibility with yeast ELP1 as coexpression of both plant genes in an elp1 elp3 yeast mutant restored Elongator's tRNA modification function in vivo. Similarly, AtELP1, not ScELP1 also supported partial complementation by yeast–plant Elp3 hybrids suggesting that AtElp1 has less stringent sequence requirements for Elp3 than ScElp1. We conclude that yeast and plant Elongator share tRNA modification roles and propose that this function might be conserved in Elongator from all eukaryotic kingdoms of life. 相似文献
93.
Endocytosis of apolipoprotein A-V by members of the low density lipoprotein receptor and the VPS10p domain receptor families 总被引:1,自引:0,他引:1
Nilsson SK Christensen S Raarup MK Ryan RO Nielsen MS Olivecrona G 《The Journal of biological chemistry》2008,283(38):25920-25927
Apolipoprotein A-V (apoA-V) is present in low amounts in plasma and has been found to modulate triacylglycerol levels in humans and in animal models. ApoA-V displays affinity for members of the low density lipoprotein receptor (LDL-R) gene family, known as the classical lipoprotein receptors, including LRP1 and SorLA/LR11. In addition to LDL-A binding repeats, the mosaic receptor SorLA/LR11 also possesses a Vps10p domain. Here we show that apoA-V also binds to sortilin, a receptor from the Vsp10p domain gene family that lacks LDL-A repeats. Binding of apoA-V to sortilin was competed by neurotensin, a ligand that binds specifically to the Vps10p domain. To investigate the biological fate of receptor-bound apoA-V, binding experiments were conducted with cultured human embryonic kidney cells transfected with either SorLA/LR11 or sortilin. Compared with nontransfected cells, apoA-V binding to SorLA/LR11- and sortilin-expressing cells was markedly enhanced. Internalization experiments, live imaging studies, and fluorescence resonance energy transfer analyses demonstrated that labeled apoA-V was rapidly internalized, co-localized with receptors in early endosomes, and followed the receptors through endosomes to the trans-Golgi network. The observed decrease of fluorescence signal intensity as a function of time during live imaging experiments suggested ligand uncoupling in endosomes with subsequent delivery to lysosomes for degradation. This interpretation was supported by experiments with (125)I-labeled apoA-V, demonstrating clear differences in degradation between transfected and nontransfected cells. We conclude that apoA-V binds to receptors possessing LDL-A repeats and Vsp10p domains and that apoA-V is internalized into cells via these receptors. This could be a mechanism by which apoA-V modulates lipoprotein metabolism in vivo. 相似文献
94.
Da Lozzo EJ Mangrich AS Rocha ME de Oliveira MB Carnieri EG 《Cell biochemistry and function》2002,20(1):19-29
The ability of the mycotoxin citrinin to act as an inhibitor of iron-induced lipoperoxidation of biological membranes prompted us to determine whether it could act as an iron chelating agent, interfering with iron redox reactions or acting as a free radical scavenger. The addition of Fe3+ to citrinin rapidly produced a chromogen, indicating the formation of citrinin-Fe3+ complexes. An EPR study confirms that citrinin acts as a ligand of Fe3+, the complexation depending on the [Fe3+]:[citrinin] ratios. Effects of citrinin on the iron redox cycle were evaluated by oxygen consumption or the o-phenanthroline test. No effect on EDTA-Fe2+-->EDTA-Fe3+ oxidation was observed in the presence of citrinin, but the mycotoxin inhibited, in a dose-dependent manner, the oxidation of Fe2+ to Fe3+ by hydrogen peroxide. Reducing agents such as ascorbic acid and DTT reduced the Fe3+-citrinin complex, but DTT did not cause reduction of Fe3+-EDTA, indicating that the redox potentials of Fe3+-citrinin and Fe3+-EDTA are not the same. The Fe2+ formed from the reduction of Fe3+-citrinin by reducing agents was not rapidly reoxidized to Fe3+ by atmospheric oxygen. Citrinin has no radical scavenger ability as demonstrated by the absence of DPPH reduction. However, a reaction between citrinin and hydrogen peroxide was observed by UV spectrum changes of citrinin after incubation with hydrogen peroxide. It was also observed that citrinin did not induce direct or reductive mobilization of iron from ferritin. These results indicate that the protective effect on iron-induced lipid peroxidation by citrinin occurs due to the formation of a redox inactive Fe3+-citrinin complex, as well as from the reaction of citrinin and hydrogen peroxide. 相似文献
95.
Previous studies have revealed that chlorine participates in a complex biogeochemical cycle in soil, which suggests that the
transport of chloride through catchments may also be influenced. The present study is based on field observations of organic
carbon, chloride (Clin), and chlorinated organic carbon (Clorg) in precipitation, soil, and runoff over a 2-year period from a small, forested catchment in southeast Sweden. The study
reveals that (1) the soil pool is dominated by Clorg, (2) the input via wet deposition and output of Clin via runoff is 30 times smaller than the total storage of chlorine (Clin + Clorg) in soil, and (3) the transport is dominated by Clin. The organic matter that entered the outlet of the catchment was more chlorinated in the autumn than during the rest of the
year, and rain events taking place in low-flow periods had a greater influence on TOC, Clorg, and Clin than did rain events taking place in high-flow periods. The seasonal pattern in combination with the low-flow versus high-flow
pattern and previous findings of increasing chlorine-to-carbon ratios with soil depth suggests that the chlorine-to-carbon
ratio variation in the leached organic matter is due that water preferentially comes from deeper layers in low-flow conditions.
This study provides well-founded estimates of Clorg and Clin storage and fluxes for the studied catchment; however, the processes underlying the observed seasonal Clorg variations and transportation processes need further study. 相似文献
96.
Navid Madani Amy M. Princiotto David Easterhoff Todd Bradley Kan Luo Wilton B. Williams Hua-Xin Liao M. Anthony Moody Ganesh E. Phad Néstor Vázquez Bernat Bruno Melillo Sampa Santra Amos B. Smith III Gunilla B. Karlsson Hedestam Barton Haynes Joseph Sodroski 《Journal of virology》2016,90(10):5031-5046
97.
Warren S. Pear Gunilla Wahlström Josiane Szpirer Göran Levan George Klein Janos Sümegi 《Immunogenetics》1986,23(6):393-395
We have previously used rat/mouse somatic cell hybrids to localize the rat c-myc gene to chromosome 7 (Sümegi et al. 1983) and the rat immunoglobulin kappa locus to chromosome 4 (Perlmann et al. 1985). We now report that by utilizing rat/mouse somatic cell hybrids, we have localized the rat immunoglobulin heavy chain locus to chromosome 6. 相似文献
98.
Medium conditioned by embryonic chick heart cells is known to support extensive neurite outgrowth from autonomic and sensory neurons. In the present report we describe the use of microcarrier cell culture with serum-free media to scale up the production of the nerve growth-stimulating factors. A growth medium composed of DME /F10 supplemented with insulin, transferrin, human serum albumin and fibronectin in combination with a low molecular weight (MW) fraction of fetal calf serum (FCS) or a mixture of FGF, dexamethasone, calmodulin and thrombin supported the heart cell proliferation at a rate similar to that of medium with 10% FCS. Furthermore, the level of successively accumulated nerve growth activity measured in a bioassay with sympathetic ganglia proved to be nearly equivalent to what was obtained when cells were grown in medium containing serum. The results confirm the potential of microcarrier cell culture in serum-free media for the production and subsequent recovery of a specific cell product. 相似文献
99.
This study examines nest building and mating in relation to size in male corkwing wrasse, Symphodus melops, in the absence of male–male competition. We kept males of varying size together with mature females in tanks containing adequate materials and sites for nest construction. Our results indicate that the size of territorial males does not influence the ability to construct a nest. However, larger males initiated nest building earlier than smaller males and nest size correlated positively with male size. Furthermore, male size did not affect whether or not a nesting male succeeded in mating, indicating that male size was not essential for female acceptance (i.e. whether or not a female is willing to mate) of a male in the current situation. 相似文献
100.