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991.
Fernández-Trillo J Barros F Machín A Carretero L Domínguez P de la Peña P 《PloS one》2011,6(9):e24674
A conserved eag domain in the cytoplasmic amino terminus of the human ether-a-go-go-related gene (hERG) potassium channel is critical for its slow deactivation gating. Introduction of gene fragments encoding the eag domain are able to restore normal deactivation properties of channels from which most of the amino terminus has been deleted, and also those lacking exclusively the eag domain or carrying a single point mutation in the initial residues of the N-terminus. Deactivation slowing in the presence of the recombinant domain is not observed with channels carrying a specific Y542C point mutation in the S4-S5 linker. On the other hand, mutations in some initial positions of the recombinant fragment also impair its ability to restore normal deactivation. Fluorescence resonance energy transfer (FRET) analysis of fluorophore-tagged proteins under total internal reflection fluorescence (TIRF) conditions revealed a substantial level of FRET between the introduced N-terminal eag fragments and the eag domain-deleted channels expressed at the membrane, but not between the recombinant eag domain and full-length channels with an intact amino terminus. The FRET signals were also minimized when the recombinant eag fragments carried single point mutations in the initial portion of their amino end, and when Y542C mutated channels were used. These data suggest that the restoration of normal deactivation gating by the N-terminal recombinant eag fragment is an intrinsic effect of this domain directed by the interaction of its N-terminal segment with the gating machinery, likely at the level of the S4-S5 linker. 相似文献
992.
Transcription of the NR1 subunit of the N-methyl-D-aspartate receptor is down-regulated by excitotoxic stimulation and cerebral ischemia 总被引:3,自引:0,他引:3
Gascón S Deogracias R Sobrado M Roda JM Renart J Rodríguez-Peña A Díaz-Guerra M 《The Journal of biological chemistry》2005,280(41):35018-35027
The N-methyl-D-aspartate (NMDA) type of glutamate receptor (NMDAR) plays central roles in normal and pathological neuronal functioning. We have examined the regulation of the NR1 subunit of the NMDAR in response to excessive activation of this receptor in in vitro and in vivo models of excitotoxicity. NR1 protein expression in cultured cortical neurons was specifically reduced by stimulation with 100 microM NMDA or glutamate. NMDA decreased NR1 protein amounts by 71% after 8 h. Low NMDA concentrations (< or = 10 microM) had no effect. NR1 down-regulation was inhibited by the general NMDAR antagonist DL-AP5 and also by ifenprodil, which specifically antagonizes NMDARs containing NR2B subunits. Arrest of NMDAR signaling with DL-AP5 after brief exposure to NMDA did not prevent subsequent NR1 decrease. Down-regulation of NR1 did not involve calpain cleavage but resulted from a decrease in de novo synthesis consequence of reduced mRNA amounts. In contrast, NMDA did not alter the expression of NR2A mRNA or newly synthesized protein. In neurons transiently transfected with an NR1 promoter/luciferase reporter construct, promoter activity was reduced by 68% after 2 h of stimulation with NMDA, and its inhibition required extracellular calcium. A similar mechanism of autoregulation of the receptor probably operates during cerebral ischemia, because NR1 mRNA and protein were strongly decreased at early stages of blood reperfusion in the infarcted brains of rats subjected to occlusion of the middle cerebral artery. Because NR1 is the obligatory subunit of NMDARs, this regulatory mechanism will be fundamental to NMDAR functioning. 相似文献
993.
Belén Chaves-Arquero David Pantoja-Uceda Alicia Roque Inmaculada Ponte Pedro Suau M. Angeles Jiménez 《Journal of biomolecular NMR》2018,72(3-4):139-148
The C-terminal domain of histone H1.0 (C-H1.0) is involved in DNA binding and is a main determinant of the chromatin condensing properties of histone H1.0. Phosphorylation at the (S/T)-P-X-(K/R) motifs affects DNA binding and is crucial for regulation of C-H1.0 function. Since C-H1.0 is an intrinsically disordered domain, solution NMR is an excellent approach to characterize the effect of phosphorylation on the structural and dynamic properties of C-H1.0. However, its very repetitive, low-amino acid-diverse and Pro-rich sequence, together with the low signal dispersion observed at the 1H–15N HSQC spectra of both non- and tri-phosphorylated C-H1.0 preclude the use of standard 1H-detected assignment strategies. We have achieved an essentially complete assignment of the heavy backbone atoms (15N, 13C′ and 13Cα), as well as 1HN and 13Cβ nuclei, of non- and tri-phosphorylated C-H1.0 by applying a novel 13C-detected CON-based strategy. No C-H1.0 region with a clear secondary structure tendency was detected by chemical shift analyses, confirming at residue level that C-H1.0 is disordered in aqueous solution. Phosphorylation only affected the chemical shifts of phosphorylated Thr’s, and their adjacent residues. Heteronuclear {1H}–15N NOEs were also essentially equal in the non- and tri-phosphorylated states. Hence, structural tendencies and dynamic properties of C-H1.0 free in aqueous solution are unmodified by phosphorylation. We propose that the assignment strategy used for C-H1.0, which is based on the acquisition of only a few 3D spectra, is an excellent choice for short-lived intrinsically disordered proteins with repetitive sequences. 相似文献
994.
Th1/Th2 cytokine balance and nitric oxide in cerebrospinal fluid and serum from patients with multiple sclerosis 总被引:3,自引:0,他引:3
Rodríguez-Sáinz Mdel C Sánchez-Ramón S de Andrés C Rodríguez-Mahou M Muñoz-Fernández MA 《European cytokine network》2002,13(1):110-114
Tumor necrosis factor (TNF-alpha) and IL-10 are key regulators of the T helper (Th)1/Th2 balance, which is critically skewed in many pathological conditions including immune-mediated inflammatory diseases of central nervous system (CNS) such as multiple sclerosis (MS). Nitric oxide (NO) has been reported to have dual effects on CNS pathology, and to play an important role in MS. We performed a cross-sectional study in 17 randomly selected patients during MS flare-up, and compared levels of TNF-alpha, IL-10 and NO in serum and cerebrospinal fluid (CSF) with the serum values of these mediators in two different control groups, healthy subjects and HIV-infected untreated patients. Serum and CSF values of TNF-alpha, IL-10 and NO were higher in MS patients than in the serum of healthy controls. Two MS patients showed increased levels of NO in CSF, with inversion of the NO(SERUM)/NO(CSF) quotient, which is clearly indicative of an intrathecal production of NO. No correlation among the values of both cytokines and NO, and the laboratory parameters analysed in MS patients (IgG index, presence of IgG oligoclonal bands and albumin quotient) was found. The high levels of TNF-alpha and IL-10 (both in serum and CSF) accompanying an MS attack suggest a simultaneous expression of Th1 and Th2 cytokines as opposed to sequential expression of Th1 followed by Th2 as described in the models of experimental autoimmune encephalomyelitis (EAE). Globally, our results support the inherent heterogeneity of the disease. 相似文献
995.
Angeles Manresa Maria Josè Espuny Jesus Guinea Francisco Comelles 《Applied microbiology and biotechnology》1987,26(4):347-351
Summary A new strain, Pseudomonas sp. GSP-910 has been isolated from soil and has been found to produce large quantities of an extracellular, highly viscous polysaccharide in a simple salt medium. Good polymer production (6.16 g·l-1) occurs on a sucrose-containing medium (2%) at high phosphate concentration (80 mM·l-1) and 0.5 g·l-1 of nitrogen source NH4Cl. The relative proportions of sugars in the polymer are: glucuronic acid 8.8%, glucose 28.07%, galactose 56.8%, and it is partially acetylated (6.32%). The isolated polymer exhibits higher viscosity at dilute concentrations than xanthan gum and it is stable at different temperatures, over a wide range of pH and in the presence of monovalent salt. In the presence of divalent cation (CaCl2 0.5%), 910-gum in aqueous solution (1%) solidifies to a resilient gel. 相似文献
996.
Aires CC Soveral G Luís PB ten Brink HJ de Almeida IT Duran M Wanders RJ Silva MF 《FEBS letters》2008,582(23-24):3359-3366
The pyruvate uptake rate in inverted submitochondrial vesicles prepared from rat liver was optimized and further characterized; the potential inhibitory effects of the anticonvulsive drug valproic acid or 2-n-propyl-pentanoic acid (VPA), Delta4-valproic acid or 2-n-propyl-4-pentenoic acid and the respective coenzyme A (CoA) conjugates were studied in the presence of a proton gradient. All tested VPA metabolites inhibited the pyruvate uptake, but the CoA esters were stronger inhibitors (40% and 60% inhibition, respectively, for valproyl-CoA and Delta4-valproyl-CoA, at 1mM). At the same concentration, the specific inhibitor 2-cyano-4-hydroxycinnamate decreased the pyruvate uptake rate by 70%. The reported inhibition of the mitochondrial pyruvate uptake may explain the significant impairment of the pyruvate-driven oxidative phosphorylation induced by VPA. 相似文献
997.
Ngoc-Loi Nguyen Joanna Pawłowska Inès Barrenechea Angeles Marek Zajaczkowski Jan Pawłowski 《Geobiology》2023,21(1):133-150
Arctic marine biodiversity is undergoing rapid changes due to global warming and modifications of oceanic water masses circulation. These changes have been demonstrated in the case of mega- and macrofauna, but much less is known about their impact on the biodiversity of smaller size organisms, such as foraminifera that represent a main component of meiofauna in the Arctic. Several studies analyzed the distribution and diversity of Arctic foraminifera. However, all these studies are based exclusively on the morphological identification of specimens sorted from sediment samples. Here, we present the first assessment of Arctic foraminifera diversity based on metabarcoding of sediment DNA samples collected in fjords and open sea areas in the Svalbard Archipelago. We obtained a total of 5,968,786 reads that represented 1384 amplicon sequence variants (ASVs). More than half of the ASVs (51.7%) could not be assigned to any group in the reference database suggesting a high genetic novelty of Svalbard foraminifera. The sieved and unsieved samples resolved comparable communities, sharing 1023 ASVs, comprising over 97% of reads. Our analyses show that the foraminiferal assemblage differs between the localities, with communities distinctly separated between fjord and open sea stations. Each locality was characterized by a specific assemblage, with only a small overlap in the case of open sea areas. Our study demonstrates a clear pattern of the influence of water masses on the structure of foraminiferal communities. The stations situated on the western coast of Svalbard that are strongly influenced by warm and salty Atlantic water (AW) are characterized by much higher diversity than stations in the northern and eastern part, where the impact of AW is less pronounced. This high diversity and specificity of Svalbard foraminifera associated with water mass distribution indicate that the foraminiferal metabarcoding data can be very useful for inferring present and past environmental conditions in the Arctic. 相似文献
998.
Liliana Rojo Fernando García-Carreño Maria de los Angeles Navarrete del Toro 《Marine biotechnology (New York, N.Y.)》2013,15(1):87-96
Aspartic proteinases in the gastric fluid of clawed lobsters Homarus americanus and Homarus gammarus were isolated to homogeneity by single-step pepstatin-A affinity chromatography; such enzymes have been previously identified as cathepsin D-like enzymes based on their deduced amino acid sequence. Here, we describe their biochemical characteristics; the properties of the lobster enzymes were compared with those of its homolog, bovine cathepsin D, and found to be unique in a number of ways. The lobster enzymes demonstrated hydrolytic activity against synthetic and natural substrates at a wider range of pH; they were more temperature-sensitive, showed no changes in the K M value at 4°C, 10°C, and 25°C, and had 20-fold higher k cat /K M values than bovine enzyme. The bovine enzyme was temperature-dependent. We propose that both properties arose from an increase in molecular flexibility required to compensate for the reduction of reaction rates at low habitat temperatures. This is supported by the fast denaturation rates induced by temperature. 相似文献
999.
1000.
Aim Data from packrat middens have established a hypothesized historical biogeography of piñon pine, Pinus edulis, including locations of glacial refugia in the south‐western USA and subsequent migration out of the refugia. In this study, we used molecular techniques to test the glacial refugial hypotheses inferred from packrat (Neotoma) midden data for P. edulis. Location South‐western USA. Methods Two fragments of chloroplast DNA (a portion of the matK gene and a portion of the rbcL gene) for a total of 1045 base pairs were amplified and sequenced for 100 individuals. Thirty‐one populations were sampled throughout the range of P. edulis. Phylogenetic analyses included maximum parsimony and maximum likelihood. Results Very little variation existed among the individuals sampled. Four haplotypes were identified. The inferred ancestral haplotype was the most widespread; it was most common in Texas and New Mexico where, with the exception of one individual, it was the only haplotype found. Arizona and Utah populations were more diverse, with almost half of the populations containing two or more haplotypes. The most derived haplotype was most abundant in Arizona. Main conclusions The distribution of haplotypes is geographically informative. Only one haplotype exists in the south‐eastern portion of the range of P. edulis whereas up to four haplotypes are found in other populations, suggesting one of two hypotheses: either all modern populations are descended from a refugial population in central Arizona, or modern populations are descended from two refugial populations, one in central Arizona and another in Texas–southern New Mexico. Interpreting these data in the light of packrat midden data gives more support for the latter hypothesis. 相似文献