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41.
Glutamine 105 in the third transmembrane helix of the thyrotropin-releasing hormone receptor (TRH-R) occupies a position equivalent to a conserved negatively charged residue in receptors for biogenic amines where it acts as counterion interacting with the cationic amine moiety of the ligand. Maximum levels of response to TRH in oocytes expressing wild-type TRH-Rs were indistinguishable from those of oocytes expressing receptors mutated to Glu, Asn, or Asp in position 105. However, the EC50 values for activation of oocyte responses increased more than 500 times in oocytes expressing mutant Glu105 receptors, in which the amido group of Gln105 has been removed by site-directed mutagenesis. Charge effects do not seem to be involved in the huge effect of mutating Gln105 to Glu, since mutation of Gln105 to Asp induces only a 15-fold increase in EC50. Furthermore, no change in EC50 is observed after mutation of Asn110 to Asp. The affinity shift (identified by changes in EC50 values for systems of comparable efficacy) in Glu105 mutant receptors was partially recovered in oocytes expressing Asn105 mutant receptors. These results and those obtained after substitution of Lys, Leu, Tyr, and Ser for Gln105 suggest that the presence and the correct position of the Gln hydrogen bond-donor amido group are important for normal functionality of the receptor. In wild type or Asp105 mutant receptors showing the same maximal responses, decreases in affinity with TRH and methyl-histidyl-TRH correlated with increased dissociation rates of hormone from the receptor. Rapid dilution experiments following subsecond stimulation indicate that the TRH-R is converted rapidly from a form showing fast dissociation kinetics to a form from which the hormone dissociates slowly. Mutation of residue 105 impairs the receptor shift between these two forms. This effect was demonstrated in a direct way by comparing [3H]methyl-histidyl-TRH dissociation rates in COS-7 cells transfected with either wild type or Asp105 mutant TRH-Rs. Thus, residues located in transmembrane helix III positions equivalent to those of the counterions for biogenic amines, regulate hormone-receptor interactions in the TRH receptor (and perhaps other receptors). Furthermore, the nature of the amino acid in these positions may also play a role, directly or indirectly, in conformational changes leading to receptor activation, and hence to signal transduction. 相似文献
42.
Summary Polymeric antioxidants prepared from 2,6-di-tertiary-butyl-4-vinyl (4-lsopropenyl)phenol and butadiene or isoprene, and their hydrogenated products (6-8 mol % of phenolic AO in the polymer) were tested by oxygen uptake studies for their effectiveness as antioxidants for polyolefins and polydienes. The polymeric antioxidants seem to be slightly less effective in short-term protection against oxidation at 150° C as compared to low molecular weight antioxidants, but more effective in long-term protection of the polymer samples at a level of 0.1 weight percent.Part 43: P. Grosso, O. Vogl: J. Macromol. Sci. Chem., in pressNow Chemical Company, Midland, MI 48674, USATo whom all correspondence should be addressed: Herman F. Mark Professor of Polymer Science, Polytechnic Institute of New York, Brooklyn, NY 11201, USA 相似文献
43.
RJ Rottier CN Hahn LW Mann M del Pilar Martin RJ Smeyne K Suzuki A d'Azzo 《Canadian Metallurgical Quarterly》1998,7(11):1787-1794
Protective protein/cathepsin A (PPCA) is a pleiotropic lysosomal enzyme that complexes with beta-galactosidase and neuraminidase, and possesses serine carboxypeptidase activity. Its deficiency in man results in the neurodegenerative lysosomal storage disorder galactosialidosis (GS). The mouse model of this disease resembles the human early onset phenotype and results in severe nephropathy and ataxia. To understand better the pathophysiology of the disease, we compared the occurrence of lysosomal PPCA mRNA and protein in normal adult mouse tissues with the incidence of lysosomal storage in PPCA(-/-) mice. PPCA expression was markedly variable among different tissues. Most sites that produced both mRNA and protein at high levels in normal mice showed extensive and overt storage in the knockout mice. However, this correlation was not consistent as some cells that normally expressed high levels of PPCA were unaffected in their storage capability in the PPCA(-/-) mice. In addition, some normally low expressing cells accumulated large amounts of undegraded products in the GS mouse. This apparent discrepancy may reflect a requirement for the catalytic rather than the protective function of PPCA and/or the presence of cell-specific substrates in certain cell types. A detailed map showing the cellular distribution of PPCA in nomal mouse tissues as well as the sites of lysosomal storage in deficient mice is critical for accurate assessment of the effects of therapeutic interventions. 相似文献
44.
45.
A new application is proposed for the on-line coupling of reversed-phase liquid chromatography to gas chromatography (RPLC-GC) that allows the GC chirospecific analysis of gamma-lactones in fruits and commercially available fruit-containing products. The use of a programmed temperature vaporizer as an interface with the system makes the transfer of large volume fractions (i.e., 2520 microL) of aqueous eluents from LC to GC possible (speed of sample transfer, 1800 microL/min). Relative standard deviations obtained for the investigated lactones under the experimental conditions vary from 7 to 14%. The described system enlarges the LC-GC application field and overcomes the limitations reported thus far concerning the use of typical normal-phase eluents (i.e., the transfer of rather small volume fractions at low speeds of sample introduction). 相似文献
46.
47.
Carrascal Ginés del Barrio Alberto A. Botella Guillermo 《The Journal of supercomputing》2021,77(3):2770-2799
The Journal of Supercomputing - Quantum computing is a reality that presents challenges to computer engineering students and practitioners. It has been claimed that it is possible to effectively... 相似文献
48.
del Campo I Basterretxea K Echanobe J Bosque G Doctor F 《IEEE transactions on systems, man, and cybernetics. Part B, Cybernetics》2012,42(2):501-512
This paper presents the development of a neuro-fuzzy agent for ambient-intelligence environments. The agent has been implemented as a system-on-chip (SoC) on a reconfigurable device, i.e., a field-programmable gate array. It is a hardware/software (HW/SW) architecture developed around a MicroBlaze processor (SW partition) and a set of parallel intellectual property cores for neuro-fuzzy modeling (HW partition). The SoC is an autonomous electronic device able to perform real-time control of the environment in a personalized and adaptive way, anticipating the desires and needs of its inhabitants. The scheme used to model the intelligent agent is a particular class of an adaptive neuro-fuzzy inference system with piecewise multilinear behavior. The main characteristics of our model are computational efficiency, scalability, and universal approximation capability. Several online experiments have been performed with data obtained in a real ubiquitous computing environment test bed. Results obtained show that the SoC is able to provide high-performance control and adaptation in a life-long mode while retaining the modeling capabilities of similar agent-based approaches implemented on larger computing machines. 相似文献
49.
López-Castejón Maria Luisa Hurtado Maria del Carmen de la Fuente Julia Mena Baltasar Bengoechea Carlos 《Iranian Polymer Journal》2021,30(7):723-735
Iranian Polymer Journal - The present work focuses on the assessment of the ability of porcine plasma protein (PPP) to be electrospun satisfactorily to form fibre mats, and their rheological and... 相似文献
50.
Davide Barbagallo Angela Caponnetto Cristina Barbagallo Rosalia Battaglia Federica Mirabella Duilia Brex Michele Stella Giuseppe Broggi Roberto Altieri Francesco Certo Rosario Caltabiano Giuseppe Maria Vincenzo Barbagallo Carmelina Daniela Anfuso Gabriella Lupo Marco Ragusa Cinzia Di Pietro Thomas Birkballe Hansen Michele Purrello 《International journal of molecular sciences》2021,22(4)
Circular RNAs (circRNAs) are a large class of RNAs with regulatory functions within cells. We recently showed that circSMARCA5 is a tumor suppressor in glioblastoma multiforme (GBM) and acts as a decoy for Serine and Arginine Rich Splicing Factor 1 (SRSF1) through six predicted binding sites (BSs). Here we characterized RNA motifs functionally involved in the interaction between circSMARCA5 and SRSF1. Three different circSMARCA5 molecules (Mut1, Mut2, Mut3), each mutated in two predicted SRSF1 BSs at once, were obtained through PCR-based replacement of wild-type (WT) BS sequences and cloned in three independent pcDNA3 vectors. Mut1 significantly decreased its capability to interact with SRSF1 as compared to WT, based on the RNA immunoprecipitation assay. In silico analysis through the “Find Individual Motif Occurrences” (FIMO) algorithm showed GAUGAA as an experimentally validated SRSF1 binding motif significantly overrepresented within both predicted SRSF1 BSs mutated in Mut1 (q-value = 0.0011). U87MG and CAS-1, transfected with Mut1, significantly increased their migration with respect to controls transfected with WT, as revealed by the cell exclusion zone assay. Immortalized human brain microvascular endothelial cells (IM-HBMEC) exposed to conditioned medium (CM) harvested from U87MG and CAS-1 transfected with Mut1 significantly sprouted more than those treated with CM harvested from U87MG and CAS-1 transfected with WT, as shown by the tube formation assay. qRT-PCR showed that the intracellular pro- to anti-angiogenic Vascular Endothelial Growth Factor A (VEGFA) mRNA isoform ratio and the amount of total VEGFA mRNA secreted in CM significantly increased in Mut1-transfected CAS-1 as compared to controls transfected with WT. Our data suggest that GAUGAA is the RNA motif responsible for the interaction between circSMARCA5 and SRSF1 as well as for the circSMARCA5-mediated control of GBM cell migration and angiogenic potential. 相似文献