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41.
A versatile strategy to define the phosphorylation preferences of plant protein kinases and screen for putative substrates 总被引:1,自引:0,他引:1
Vlad F Turk BE Peynot P Leung J Merlot S 《The Plant journal : for cell and molecular biology》2008,55(1):104-117
Most signaling networks are regulated by reversible protein phosphorylation. The specificity of this regulation depends in part on the capacity of protein kinases to recognize and efficiently phosphorylate particular sequence motifs in their substrates. Sequenced plant genomes potentially encode over than 1000 protein kinases, representing 4% of the proteins, twice the proportion found in humans. This plethora of plant kinases requires the development of high-throughput strategies to identify their substrates. In this study, we have implemented a semi-degenerate peptide array screen to define the phosphorylation preferences of four kinases from Arabidopsis thaliana that are representative of the plant calcium-dependent protein kinase and Snf1-related kinase superfamily. We converted these quantitative data into position-specific scoring matrices to identify putative substrates of these kinases in silico in protein sequence databases. Our data show that these kinases display related but nevertheless distinct phosphorylation motif preferences, suggesting that they might share common targets but are likely to have specific substrates. Our analysis also reveals that a conserved motif found in the stress-related dehydrin protein family may be targeted by the SnRK2-10 kinase. Our results indicate that semi-degenerate peptide array screening is a versatile strategy that can be used on numerous plant kinases to facilitate identification of their substrates, and therefore represents a valuable tool to decipher phosphorylation-regulated signaling networks in plants. 相似文献
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Letellier K Azeddine B Blain S Turgeon I Wang da S Boiro MS Moldovan F Labelle H Poitras B Rivard CH Grimard G Parent S Ouellet J Lacroix G Moreau A 《Médecine sciences : M/S》2007,23(11):910-916
Adolescent idiopathic scoliosis (AIS) is the most common form of scoliosis that affects a significant number of young teenagers, mainly females (0.2-6 % of the population). Historically, several hypothesis were postulated to explain the aetiology of AIS, including genetic factors, biochemical factors, mechanics, neurological, muscular factors and hormonal factors. The neuroendocrine hypothesis involving a melatonin deficiency as the source for AIS has generated great interest. This hypothesis stems from the fact that experimental pinealectomy in chicken, and more recently in rats maintained in a bipedal mode, produces a scoliosis. The biological relevance of melatonin in idiopathic scoliosis is controversial since no significant decrease in circulating melatonin level has been observed in a majority of studies. Analysis of melatonin signal transduction in musculoskeletal tissues of AIS patients demonstrated for the first time a defect occurring in a cell autonomous manner in different cell types isolated from AIS patients suffering of the most severe form of that disease. These results have led to a classification of AIS patients in three different functional groups depending on their response to melatonin, suggesting that the cause of AIS involves several genes. Molecular analysis showed that melatonin signaling dysfunction is triggered by an increased phosphorylation of Gi proteins inactivating their function. This discovery has led to development of a first scoliosis screening assay. This test, using blood sample, is currently in clinical validation process in Canada and could be used for screening children at high risk of developing AIS. 相似文献
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Jiri Soukup Helena Hornychova Monika Manethova Kvetoslava Michalova Ludmila Michnova Lenka Popovska Veronika Skarkova Tomas Cesak David Netuka Ales Ryska Jan Cap Václav Hána Václav Hána Jr Michal Kršek Eva Dvořáková Michal Krčma Ivica Lazurova Věra Olšovská Karel Starý Peter Vaňuga Filip Gabalec 《Journal of cellular and molecular medicine》2021,25(5):2484-2492
In somatotroph pituitary tumours, somatostatin analogue (SSA) therapy outcomes vary throughout the studies. We performed an analysis of cohort of patients with acromegaly from the Czech registry to identify new prognostic and predictive factors. Clinical data of patients were collected, and complex immunohistochemical assessment of tumour samples was performed (SSTR1-5, dopamine D2 receptor, E-cadherin, AIP). The study included 110 patients. In 31, SSA treatment outcome was evaluated. Sparsely granulated tumours (SGST) differed from the other subtypes in expression of SSTR2A, SSTR3, SSTR5 and E-cadherin and occurred more often in young. No other clinical differences were observed. Trouillas grading system showed association with age, tumour size and SSTR2A expression. Factors significantly associated with SSA treatment outcome included age, IGF1 levels, tumour size and expression of E-cadherin and SSTR2A. In the group of SGST, poor SSA response was observed in younger patients with larger tumours, lower levels of SSTR2A and higher Ki67. We observed no relationship with expression of other proteins including AIP. No predictive value of E-cadherin was observed when tumour subtype was considered . Multiple additional factors apart from SSTR2A expression can predict treatment outcome in patients with acromegaly. 相似文献
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Simon Ströbel Marko Loparic David Wendt Andreas D Schenk Christian Candrian Raija LP Lindberg Florina Moldovan Andrea Barbero Ivan Martin 《Arthritis research & therapy》2010,12(2):R34
Introduction
Oxygen is a critical parameter proposed to modulate the functions of chondrocytes ex-vivo as well as in damaged joints. This article investigates the effect of low (more physiological) oxygen percentage on the biosynthetic and catabolic activity of human articular chondrocytes (HAC) at different phases of in vitro culture. 相似文献48.
Deac F Iacob B Fischer-Fodor E Damian G Silaghi-Dumitrescu R 《Journal of biochemistry》2011,149(1):75-82
Periodate modification of the sugar moiety in sugars, including adenosine triphosphate (ATP), has previously been employed in order to prepare dialdehyde-type reagents, which were then utilized in crosslinking reactions on haemoglobin, yielding polymerized material with useful dioxygen-binding properties and hence proposed as possible artificial oxygen carriers ('blood substitutes'). Here, the periodate protocol is shown to be applicable to a wider range of oxygen-containing compounds, illustrated by starch and polyethylene glycol. Derivatization protocols are described for haemoglobin with such periodate-treated crosslinking agents, and the dioxygen-binding properties and redox reactivities are investigated for the derivatized haemoglobins, with emphasis on pro-oxidative properties. There is a general tendency of the derivatization to result in higher autooxidation rates. The peroxide reactivity of the met (ferric) form is also affected by derivatization, as witnessed, among others, by varying yields of ferryl [Fe (IV)-oxo] and free radical generated. In cell, culture tests (human umbilical vein epithelial cells, HUVEC), the derivatization protocols show no toxic effect. 相似文献
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Fischer-Fodor E Mot A Deac F Arkosi M Silaghi-Dumitrescu R 《Journal of biosciences》2011,36(2):215-221
Hemerythrin is a dioxygen-carrying protein whose oxidative/nitrosative stress-related reactivity is lower than that of hemoglobin,
which may warrant investigation of hemerythrin as raw material for artificial oxygen carriers (‘blood substitutes’). We report
here the first biological tests for hemerythrin and its chemical derivatives, comparing their performance with that of a representative
competitor, glutaraldehyde-polymerized bovine hemoglobin. Hemerythrin (native or derivatized) exhibits a proliferative effect
on human umbilical vein endothelial cell (HUVEC) cultures, as opposed to a slight inhibitory effect of hemoglobin. A similar
positive effect is displayed on human lymphocytes by glutaraldehyde-polymerized hemerythrin, but not by native or polyethylene
glycol-derivatized hemerythrin. 相似文献
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The aim of the study was to evaluate the antifungal activity of a new triazole formulation against 182 clinical isolates of
yeasts recovered from blood cultures in three tertiary hospitals in Romania and to compare its activity with those of fluconazole
and voriconazole. In vitro susceptibility was assessed by following the guidelines of AFST-EUCAST E. Def. 7.1. The distribution
of minimum inhibitory concentrations (MICs) of MXP-4509 was very similar to that of voriconazole (MIC50: 0.0312 mg/l vs. 0.0156 mg/l; MIC90: 0.25 mg/l vs. 0.25 mg/l), but significantly different from that of fluconazole (MIC50: 0.0312 mg/l vs. 0.5 mg/l; MIC90: 0.25 mg/l vs. 32 mg/l). The new triazole MXP-4509 proved to have a good in vitro antifungal activity raising the interest
for further pharmacological and microbiological investigations in order to assess its potential advantages for therapy. 相似文献