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21.
Background
Mutations linked to early onset, familial forms of Alzheimer''s disease (FAD) are found most frequently in PSEN1, the gene encoding presenilin-1 (PS1). Together with nicastrin (NCT), anterior pharynx-defective protein 1 (APH1), and presenilin enhancer 2 (PEN2), the catalytic subunit PS1 constitutes the core of the γ-secretase complex and contributes to the proteolysis of the amyloid precursor protein (APP) into amyloid-beta (Aβ) peptides. Although there is a growing consensus that FAD-linked PS1 mutations affect Aβ production by enhancing the Aβ1–42/Aβ1–40 ratio, it remains unclear whether and how they affect the generation of APP intracellular domain (AICD). Moreover, controversy exists as to how PS1 mutations exert their effects in different experimental systems, by either increasing Aβ1–42 production, decreasing Aβ1–40 production, or both. Because it could be explained by the heterogeneity in the composition of γ-secretase, we purified to homogeneity complexes made of human NCT, APH1aL, PEN2, and the pathogenic PS1 mutants L166P, ΔE9, or P436Q.Methodology/Principal Findings
We took advantage of a mouse embryonic fibroblast cell line lacking PS1 and PS2 to generate different stable cell lines overexpressing human γ-secretase complexes with different FAD-linked PS1 mutations. A multi-step affinity purification procedure was used to isolate semi-purified or highly purified γ-secretase complexes. The functional characterization of these complexes revealed that all PS1 FAD-linked mutations caused a loss of γ-secretase activity phenotype, in terms of Aβ1–40, Aβ1–42 and APP intracellular domain productions in vitro.Conclusion/Significance
Our data support the view that PS1 mutations lead to a strong γ-secretase loss-of-function phenotype and an increased Aβ1–42/Aβ1–40 ratio, two mechanisms that are potentially involved in the pathogenesis of Alzheimer''s disease. 相似文献22.
Jean-Fran?ois?ManenEmail author Olga?Sinitsyna Lorène?Aeschbach Alexander?V?Markov Arkady?Sinitsyn 《BMC plant biology》2005,5(1):23
Background
DNA extraction from plant tissues, unlike DNA isolation from mammalian tissues, remains difficult due to the presence of a rigid cell wall around the plant cells. Currently used methods inevitably require a laborious mechanical grinding step, necessary to disrupt the cell wall for the release of DNA. 相似文献23.
V. Schmid B. Aeschbach K. Agata J. Kosaka S. Reber-Müller N. Sprenger G. Eguchi 《Development genes and evolution》1995,204(7-8):465-476
Extracellular matrices (ECMs) of phylogenetically very distant organisms were tested for their ability to support cell adhesion, spreading and DNA replication in reciprocal xenograft adhesion tests. Mechanically dissociated cells of the medusa Podocoryne carnea (Cnidaria, Hydrozoa) were seeded on ECMs of polyps and medusa, and on several ECM glycoproteins or entire ECMs from vertebrates. In reciprocal experiments, cells from different vertebrate cell-lines were seeded on ECMs of polyps, medusae and also on electrophoresed and blotted extracts of both types of ECMs. The results demonstrate that medusa cells adhere and spread on polyp and medusa ECMs but do not recognize vertebrate ECMs or purified ECM glycoproteins. Vertebrate cells in contrast adhere, spread and proliferate on ECMs of polyps and medusae. The number of attached cells depends on the cell type, the type of ECM and, in certain cases, on the stage of the cell cycle. Cell adhesion experiments with pretreated ECMs of polyps and medusae, e.g. oxidation of carbohydrate residues with sodium-metaperiodate, or blocking of certain carbohydrate moieties with the lectin wheat germ agglutinin or a carbohydrate-specific monoclonal antibody, demonstrate that ECM carbohydrates are more important for cell-ECM interactions of medusa cells than for vertebrate cells. Furthermore, the experiments indicate that polyp and medusa ECMs contain different components which strongly modulate adhesion, spreading and DNA replication of vertebrate cells. 相似文献
24.
Correlations between the ascorbate-glutathione pathway and effectiveness in legume root nodules 总被引:4,自引:0,他引:4
Legume root nodules use the ascorbate-glutathione pathway to remove harmful H2 O2 . In the present study. effective and ineffective nodules from soybean and alfalfa were compared with regard to this pathway. Effective nodules had higher activity of all 4 enzymes (ascorbate peroxidase, EC 1. 11. 1. 11: monodehydroascorbate reductase, EC 1. 6. 5. 4: dehydroascorbate reductase, EC 1. 8. 5. 1: and glutathione reductase, EC 1. 6. 4. 2). The concentration of thiol tripeptides (primarily homoglutathione) was about 1 m M in effective nodules – a level 3–4-fold higher than in ineffective nodules. Effective nodules contained higher levels of NAD+ . NADP+ and NADPH. but not of NADH or ascorbate. The increased capacity for peroxide scavenging in effective nodules as compared to ineffective nodules emphasizes the important protective role that this pathway may play in processes related to nitrogen fixation. 相似文献
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26.
Simon A Hinke Manuel F Navedo Allison Ulman Jennifer L Whiting Patrick J Nygren Geng Tian Antonio J Jimenez-Caliani Lorene K Langeberg Vincenzo Cirulli Anders Tengholm Mark L Dell'Acqua L Fernando Santana John D Scott 《The EMBO journal》2012,31(20):3991-4004
Endocrine release of insulin principally controls glucose homeostasis. Nutrient-induced exocytosis of insulin granules from pancreatic β-cells involves ion channels and mobilization of Ca2+ and cyclic AMP (cAMP) signalling pathways. Whole-animal physiology, islet studies and live-β-cell imaging approaches reveal that ablation of the kinase/phosphatase anchoring protein AKAP150 impairs insulin secretion in mice. Loss of AKAP150 impacts L-type Ca2+ currents, and attenuates cytoplasmic accumulation of Ca2+ and cAMP in β-cells. Yet surprisingly AKAP150 null animals display improved glucose handling and heightened insulin sensitivity in skeletal muscle. More refined analyses of AKAP150 knock-in mice unable to anchor protein kinase A or protein phosphatase 2B uncover an unexpected observation that tethering of phosphatases to a seven-residue sequence of the anchoring protein is the predominant molecular event underlying these metabolic phenotypes. Thus anchored signalling events that facilitate insulin secretion and glucose homeostasis may be set by AKAP150 associated phosphatase activity. 相似文献
27.
Adelaide Fernandes Ana Sofia Falcão Elsa Abranches Evguenia Bekman Domingos Henrique Lorene M. Lanier Dora Brites 《Developmental neurobiology》2009,69(9):568-582
Elevated levels of serum unconjugated bilirubin (UCB) in the first weeks of life may lead to long‐term neurologic impairment. We previously reported that an early exposure of developing neurons to UCB, in conditions mimicking moderate to severe neonatal jaundice, leads to neuritic atrophy and cell death. Here, we have further analyzed the effect of UCB on nerve cell differentiation and neuronal development, addressing how UCB may affect the viability of undifferentiated neural precursor cells and their fate decisions, as well as the development of hippocampal neurons in terms of dendritic and axonal elongation and branching, the axonal growth cone morphology, and the establishment of dendritic spines and synapses. Our results indicate that UCB reduces the viability of proliferating neural precursors, decreases neurogenesis without affecting astrogliogenesis, and increases cellular dysfunction in differentiating cells. In addition, an early exposure of neurons to UCB decreases the number of dendritic and axonal branches at 3 and 9 days in vitro (DIV), and a higher number of neurons showed a smaller growth cone area. UCB‐treated neurons also reveal a decreased density of dendritic spines and synapses at 21 DIV. Such deleterious role of UCB in neuronal differentiation, development, and plasticity may compromise the performance of the brain in later life. © 2009 Wiley Periodicals, Inc. Develop Neurobiol 2009 相似文献
28.
Jennifer L. Whiting Patrick J. Nygren Brian J. Tunquist Lorene K. Langeberg Ole-Morten Seternes John D. Scott 《The Journal of biological chemistry》2015,290(32):19445-19457
The proximity of an enzyme to its substrate can influence rate and magnitude of catalysis. A-kinase anchoring protein 220 (AKAP220) is a multivalent anchoring protein that can sequester a variety of signal transduction enzymes. These include protein kinase A (PKA) and glycogen synthase kinase 3β (GSK3β). Using a combination of molecular and cellular approaches we show that GSK3β phosphorylation of Thr-1132 on AKAP220 initiates recruitment of this kinase into the enzyme scaffold. We also find that AKAP220 anchors GSK3β and its substrate β-catenin in membrane ruffles. Interestingly, GSK3β can be released from the multienzyme complex in response to PKA phosphorylation on serine 9, which suppresses GSK3β activity. The signaling scaffold may enhance this regulatory mechanism, as AKAP220 has the capacity to anchor two PKA holoenzymes. Site 1 on AKAP220 (residues 610–623) preferentially interacts with RII, whereas site 2 (residues 1633–1646) exhibits a dual specificity for RI and RII. In vitro affinity measurements revealed that site 2 on AKAP220 binds RII with ∼10-fold higher affinity than site 1. Occupancy of both R subunit binding sites on AKAP220 could provide a mechanism to amplify local cAMP responses and enable cross-talk between PKA and GSK3β. 相似文献
29.
Perino A Ghigo A Ferrero E Morello F Santulli G Baillie GS Damilano F Dunlop AJ Pawson C Walser R Levi R Altruda F Silengo L Langeberg LK Neubauer G Heymans S Lembo G Wymann MP Wetzker R Houslay MD Iaccarino G Scott JD Hirsch E 《Molecular cell》2011,42(1):84-95
Adrenergic stimulation of the heart engages cAMP and phosphoinositide second messenger signaling cascades. Cardiac phosphoinositide 3-kinase p110γ participates in these processes by sustaining β-adrenergic receptor internalization through its catalytic function and by controlling phosphodiesterase 3B (PDE3B) activity via an unknown kinase-independent mechanism. We have discovered that p110γ anchors protein kinase A (PKA) through a site in its N-terminal region. Anchored PKA activates PDE3B to enhance cAMP degradation and phosphorylates p110γ to inhibit PIP(3) production. This provides local feedback control of PIP(3) and cAMP signaling events. In congestive heart failure, p110γ is upregulated and escapes PKA-mediated inhibition, contributing to a reduction in β-adrenergic receptor density. Pharmacological inhibition of p110γ normalizes β-adrenergic receptor density and improves contractility in failing hearts. 相似文献
30.
Highlights? Involvement of formin mDia3 in stable kinetochore microtubule attachment ? The actin activity of mDia3 is not necessary for metaphase chromosome alignment ? EB1-binding by mDia3 contributes to position chromosomes at the metaphase plate ? Aurora B phosphorylates mDia3 to regulate its microtubule activity 相似文献