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In recent years, phenotypic-based screens have become increasingly popular in drug discovery. A major challenge of this approach is that it does not provide information about the mechanism of action of the hits. This has led to the development of multiple strategies for target deconvolution. Thermal proteome profiling (TPP) allows for an unbiased search of drug targets and can be applied in living cells without requiring compound labeling. TPP is based on the principle that proteins become more resistant to heat-induced unfolding when complexed with a ligand, e.g., the hit compound from a phenotypic screen. The melting proteome is also sensitive to other intracellular events, such as levels of metabolites, post-translational modifications and protein-protein interactions. In this review, we describe the principles of this approach, review the method and its developments, and discuss its current and future applications. While proteomics has generally focused on measuring relative protein concentrations, TPP provides a novel approach to gather complementary information on protein stability not present in expression datasets. Therefore, this strategy has great potential not only for drug discovery, but also for answering fundamental biological questions. 相似文献
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S Tomi? L Sesarti? J Tomasi? 《Comparative biochemistry and physiology. B, Comparative biochemistry》1989,92(4):681-684
1. 14C-labelled methyl 2,6-di-O-pivaloyl-alpha-D-glucopyranoside (1) was used as a substrate for esterases from rabbit, guinea pig, mouse, donkey, pig, horse, sheep and human sera. 2. Stepwise de-esterification of the diester substrate 1 occurred with rabbit, guinea pig and mouse serum. Data on time-course experiments and kinetic data are reported. 3. The use of donkey, pig, horse, sheep and human serum led to the migration of the 2-O-pivaloyl group in substrate 1 to the position 4- in the sugar molecule, followed by stepwise de-esterifications of both 1 and the newly formed methyl 4,6-di-O-pivaloyl-alpha-D-glucopyranoside (4). A report is given on the time-course experiments. 相似文献
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Gibberellin A3 (GA3) treatment enhances the in vitro templateactivity of chromatin in peas (Pisum sativum, var. Alaska) (Tomiet al. 1983a). Effects of GA3 on the chromatin template weresurveyed for the chromatin components, DNA-tightly bound proteins(TBPs) and loosely bound proteins (LBPs). An addition of LBPsfrom GA3-treated chromatin enhanced the template activity ofLBP-depleted chromatin. In addition, both components of theLBPs, high mobility group proteins (HMGs) and low mobility groupproteins (LMGs) were required for this enhancement. The LBPsconverted LBP-depleted chromatin into a form susceptible toDNase I. LBPs probably are at least partly responsible for GA3-enhancedtemplate activity due to their relaxing the interaction of corehistones and DNA. (Received March 12, 1983; Accepted June 16, 1983) 相似文献
377.
Soili Stenroos Tomi Laukka Seppo Huhtinen Peter Döbbeler Leena Myllys Kimmo Syrjänen Jaakko Hyvönen 《Cladistics : the international journal of the Willi Hennig Society》2010,26(3):281-300
Numerous species of microscopic fungi inhabit mosses and hepatics. They are severely overlooked and their identity and nutritional strategies are mostly unknown. Most of these bryosymbiotic fungi belong to the Ascomycota. Their fruit‐bodies are extremely small, often reduced and simply structured, which is why they cannot be reliably identified and classified by their morphological and anatomical characters. A phylogenetic hypothesis of bryosymbiotic ascomycetes is presented. New sequences of 78 samples, including 61 bryosymbionts, were produced, the total amount of terminals being 206. Of these, 202 are Ascomycetes. Sequences from the following five gene loci were used: rDNA SSU, rDNA LSU, RPB2, mitochondrial rDNA SSU, and rDNA 5.8S. The program TNT was used for tree search and support value estimation. We show that bryosymbiotic fungi occur in numerous lineages, one of which represents a newly discovered lineage among the Ascomycota and exhibits a tripartite association with cyanobacteria and sphagna. A new genus Trizodia is proposed for this basal clade. Our results demonstrate that even highly specialized life strategies can be adopted multiple times during evolution, and that in many cases bryosymbionts appear to have evolved from saprobic ancestors. © The Willi Hennig Society 2009. 相似文献
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Various techniques for generation of peptide and peptidomimetic libraries are summarized in this article. Multipin, tea bag, and split-couple-mix techniques represent the major methods used to make peptides and peptidomimetics libraries. The synthesis of these libraries were made in either discrete or mixture format. Peptides and peptidomimetics combinatorial libraries were screened to discover leads against a variety of targets. These targets, including bacteria, fungus, virus, receptors, and enzymes were used in the screening of the libraries. Discovered leads can be further optimized by combinatorial approaches. 相似文献