![]() ![]() application number PCT/US2012/037131), nonetheless, the lack of fast and reliable high-throughput methods for assessing the protective capacity of proteins considerably hinders the discovery and characterization of novel proteins with such functionality. Methods for chaperone activity testing are under patenting (e.g. Such methods are effective in pooling proteins with stress-related function, but they have limited reproducibility and inherent high error rates of false positives, and require detailed in vitro studies for verification. The current state of the art in chaperone identification and discovery combines in silico homology screens, analysis of transcriptomics data, and co-purification with other proteins. Therefore functional and mechanistic characterization is crucial. This makes proteins with chaperone function potential drug targets, due to their function in various pathological and physiological processes inside the cell. In some cardiovascular diseases, cataract, alcoholic hepatitis, cystic fibrosis, phenylketonuria, and in a range of human cancers the protein homeostasis machinery was also shown to be involved as the first line of defense against the disease, thus rendering it a molecular marker for diagnostics. Overexpression of chaperones in flies and mice so far proved effective in suppressing neurodegeneration. During unfavorable environmental conditions, infection or disease, animals rely on a wide variety of chaperones to prevent and reduce the damage caused to cellular compartments and molecules. Chaperones are involved in the molecular response to stress in plants, bacteria, and animals, and were identified as key players in host-pathogen interaction and in the formation of innate and adaptive immunity. Besides physiological processes, these helper proteins also fulfil crucial functions under unfavorable environmental conditions, such as temperature or osmotic stresses, but also in disease conditions and in response to a pathogen attack. Molecular chaperones are a diverse group of proteins that play critical roles in assisting the folding and assembly of nascent protein chains, refolding proteins, assisting protein translocation through membranes, and often facilitating protein degradation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Ĭompeting interests: The authors have declared that no competing interests exist. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.ĭata Availability: All relevant data are within the paper and its Supporting Information files.įunding: This work was supported by the Odysseus grant G.0029.12 from the Fonds Wetenschappelijk Onderzoek (FWO, ) to PT, and DK is supported by a FWO postdoctoral fellowship 1.2.734.13. Received: Accepted: AugPublished: August 26, 2016Ĭopyright: © 2016 Hristozova et al. Brodsky, University of Pittsburgh, UNITED STATES The use of this method may lead to a considerable increase in the number of experimentally verified proteins with such functions, and may also allow the dissection of their molecular mechanism for a better understanding of their function.Ĭitation: Hristozova N, Tompa P, Kovacs D (2016) A Novel Method for Assessing the Chaperone Activity of Proteins. The current state of the art does not allow the in vitro measurements of chaperone activity in a highly parallel manner with high accuracy or high reproducibility, thus we believe that the method we report will be of significant benefit in this direction. Here we propose a novel technique to test and measure the capability for protective activity of known and putative chaperones in a semi-high throughput manner on a plate reader. Their molecular functions range from stabilizing stress-susceptible molecules and membranes to assisting the refolding of stress-damaged proteins, thereby acting as protective barriers against cellular damage. Among stress protein families–molecules expressed during adverse conditions, infection, and diseases–chaperones are highly abundant. Depending on their specific function, molecular chaperones are involved in a plethora of cellular processes by playing key roles in nascent protein chain folding, transport and quality control. Protein chaperones are molecular machines which function both during homeostasis and stress conditions in all living organisms. ![]()
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