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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
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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
35.
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
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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
38.
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
39.
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
40.
Sara da Mata Joana Ferreira Inmaculada Nicols Susana Esteves Gonalo Esteves Sofia Lrias Fernanda Silva Adela Saco Daniela Cochicho Mrio Cunha Marta del Pino Jaume Ordi Ana Flix 《International journal of molecular sciences》2021,22(5)
The expression of p16 is a good surrogate of human papillomavirus (HPV) infection in HPV-associated cancers. The significance of p16 expression, HPV genotype and genera in the outcome of patients with HPV-associated cervical cancer (CC) is unclear. Our aim is to ascertain the prognostic significance of these factors. Data from 348 patients (median age: 47.5 years old) with CC, diagnosed in two referral centers, were retrospectively collected. Advanced disease (FIGO2018 IB2-IV) was present in 68% of patients. A single HPV genotype was identified in 82.8% of patients. The most common HPVs were HPV16 (69%) and HPV18 (14%). HPV genera reflected this distribution. HPV16 tumors presented at an earlier stage. P16 was negative in 18 cases (5.2%), 83.3% of which were squamous cell carcinomas. These cases occurred in older patients who tended to have advanced disease. In the univariate analysis, HPV16 (HR: 0.58; p = 0.0198), α-9 genera (HR: 0.37; p = 0.0106) and p16 overexpression (HR: 0.54; p = 0.032) were associated with better survival. HPV16 (HR: 0.63; p = 0.0174) and α-9 genera (HR: 0.57; p = 0.0286) were associated with less relapse. In the multivariate analysis, only the International Federation of Gynecology and Obstetrics (FIGO) stage retained an independent prognostic value. HPV16, α-9 genera and p16 overexpression were associated with better survival, although not as independent prognostic factors. Patients with p16-negative HPV-associated CC were older, presented with advanced disease and had worse prognosis. 相似文献