دانلود رایگان مقاله لاتین فعالیت درون جانداری از سایت الزویر
عنوان فارسی مقاله:
نظارت بی درنگ آرتمین در فعالیت نگهبانی درون جانداری با استفاده از لوسیفراز به عنوان گزارشگر درون سلولی
عنوان انگلیسی مقاله:
Real-time monitoring of artemin in vivo chaperone activity using luciferase as an intracellular reporter
سال انتشار : 2016
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مقدمه انگلیسی مقاله:
1. Introduction
The environmental changes, like temperature fluctuations can affect the native conformation of proteins. High temperatures lead to accelerated formation of protein aggregates. These effects include thermodynamic instability, hydrophobic interaction, protein diffusion and chemical reactions. Protein destabilization leads to the formation of unfolded protein intermediates and promotes aggregation process [1]. The hydrophobic residues are normally buried inside the proteins, whereas by increasing the temperature, the hydrophobic amino acids are mostly oriented to the surroundings which leads to their association to form large-scale precipitates or protein aggregates [2]. The influence of low temperature and cold shock responses were investigated in different organisms including eukaryotes, bacteria and archaea [3e5]. The effect of cold shock was observed at different levels: (1) fluidity of membrane was decreased and some membrane functions could be disturbed including active transport, protein secretion and cell signaling, (2) the secondary structure in RNA and DNA were stabilized, thereby the mRNA translation rate and the transcription efficiency were reduced, (3) some proteins folded more slowly and even inappropriate folding might occur and the ribosomal functions were disturbed [6,4] and (4) proteins could be destabilized at low temperatures, resulted in generating intermediates which promoted protein aggregation [1]. In microorganisms, it was observed that cold stress could induce the synthesis of several cold-shock proteins [7]. In some literature, the effects of chaperones were studied under cold stress conditions [8e12]. Small heat shock proteins (sHSPs) with low molecular weight (12e40 kD) are exclusively expressed in responding to various environmental changes and they can be found in all organisms including bacteria, plants and animals [13]. sHSPs have protective effects on cell viability at high temperatures or other stress conditions and it is suggested that they might have a role in forming or maintaining of the native conformation of proteins in the stress conditions. They bind to partially denatured proteins and prevent irreversible protein aggregation during stress conditions [14]. High temperature causes destabilization of proteins which leads to form folding intermediates that tend to associate with temperatureactivated sHSPs. This phenomenon leads to intercellular complexes containing unstable proteins and sHSPs [15,16]. Artemin is a thermostable protein in cyst embryo of Artemia under stress conditions, and general stress resistance of this crustacea is relevant to high regulatory expression of artemin as a stress protein [17]. Artemin consists of 24 monomers with 27 kDa molecular weight that forms an oligomer with rosette-like structure and 600 kDa molecular weight [18]. Previous reports suggested different functions for artemin including promoting resistance of mammalian cells in stress conditions and increasing bacterial cell viability affected by high temperature stress in vivo [19,20], increasing protein refolding efficiency, the ability of binding to RNA at elevated temperature and its protecting heat-induced aggregation of different protein substrates in vitro [21e23]. All observations proposed that artemin acts as an efficient molecular chaperone. According to the previous studies, it was found that artemin showed significantly higher chaperone-assisted refolding and antiaggregation efficiency compared to reported chaperones. In this study, the chaperoning behavior of artemin has been further investigated and the study has focused on the protective effects of artemin on proteins in living cells. For real-time monitoring of the artemin chaperone activity, a system was used to measure the protective potential of it in living cells (Scheme 1). The used realtime reporter system provided a rapid and simple test for molecular chaperone activity assays, in which luciferase was used as a reporter and the effect of artemin was considered under stress conditions. It should be mentioned that luciferase is a model protein used as a substrate for chaperones. Different experiments were designed by using cold and heat as stressors. In the present study, the real-time reporter system is introduced for in vivo measuring chaperone activity under the stress conditions, and we have investigated, for the first time, the protective effect for artemin under cold shock.
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کلمات کلیدی:
Real-time monitoring of artemin in vivo chaperone activity using ... https://www.infona.pl/.../bwmeta1.element.elsevier-8e93e8c2-7759-3e4b-adc7-67aeee6... by Z Takalloo - 2016 - Cited by 1 - Related articles Here, co-transformation of E. coli was performed with two expression vectors ... of artemin in vivo chaperone activity using luciferase as an intracellular reporter. SID.ir | CO-EXPRESSION OF ARTEMIN WITH FIREFLY LUCIFERASE ... en.journals.sid.ir/ViewPaper.aspx?ID=473951 by Z TAKALU - 2014 Download Free Full-Text of an article CO-EXPRESSION OF ARTEMIN WITH FIREFLY ... AGGREGATION BY ARTEMIN AND ITS CHAPERONE ACTIVITY IN VIVO. Molecular Aspects of the Stress Response: Chaperones, Membranes and ... https://books.google.com/books?isbn=0387399755 Peter Csermely, László Vígh - 2007 - Science Note that only four shsps share a chaperone activity Gene HspB Other ... Extinction of thermosensitive luciferase light emission was used as an intracellular marker ... Taken together, these phenomena provide the cell with a survival advantage ... Stress Response: Methods and Protocols https://books.google.com/books?isbn=1592590543 Stephen M. Keyse - 2000 - Medical If desirable, one can check using light microscopy whether the pellets ... IN VIVO The use of foreign reporter proteins to assay chaperone activity in vivo was ... the intracellular localization of the luciferase, and the concentration of luciferase per ... Cell Stress Proteins - Page 188 - Google Books Result https://books.google.com/books?isbn=0387397175 Stuart K. Calderwood - 2009 - Science Studies using various deletion mutants showed that neither the ATP binding domain ... and the β-sheet domain prevented the heat-induced aggregation of luciferase. ... That LH and H mutants possess equivalent chaperoning activity suggests that ... Interaction of hsp110 and grp170 with Other Chaperones Intracellular 188 ...
