Design and Synthesis of a Tag‐Free Chemical Probe for Photoaffinity Labeling

Mayer, Timo; Maier, Martin E.

doi:10.1002/ejoc.200700188

Cited by 50 publications

(34 citation statements)

References 59 publications

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“…The ether fractions were collected, dried over Na 2 SO 4 , and evaporated in vacuo to afford the title compound as a clear oil (6.7 g, 64%). The 1 H proton and 13 C NMR shifts were confirmed with the report by Mayer and Maier [32].…”

Section: Methodssupporting

confidence: 90%

Synthesis and properties of oligonucleotides that contain a triazole‐linked nucleic acid dimer

Efthymiou¹,

Desaulniers²

2011

Journal of Heterocyclic Chem

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in Wiley Online Library (wileyonlinelibrary.com).New chemically modified oligonucleotides at the site of the backbone are needed to improve the properties of oligonucleotides. A practical synthesis for a triazole-linked nucleoside dimer based on a PNA-like structure has been developed. This involves synthesizing two uracil-based monomers that contain either an azide or an alkyne functionality, followed by copper-catalyzed 1,3-dipolar cycloaddition. This dimer was incorporated within an oligonucleotide via phosphoramidite chemistry and UV-monitored thermal denaturation data illustrates slight destabilization relative to its target complementary sequence. This chemically modified dimer will allow for a future investigation of its properties within DNA and RNA-based applications.

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Section: Methodssupporting

confidence: 90%

Synthesis and properties of oligonucleotides that contain a triazole‐linked nucleic acid dimer

Efthymiou¹,

Desaulniers²

2011

Journal of Heterocyclic Chem

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“…To determine whether ascr#2 binds directly to DAF-37, we used photoaffinity labeling of DAF-37 expressed in human cell culture (25,26). Chemical design of an ascr#2-derived photoaffinity-labeling probe was based on previously described methods (4,27) and incorporates functional groups that enable photo-cross-linking to the receptor and subsequent conjugation with a reporter while preserving the probe's ability to bind to DAF-37. As a photo-cross-linkable unit we chose a trifluorodiazirine, to which we linked a bio-orthogonal terminal alkyne, which can be conjugated to appropriate azide reporters via click chemistry (Fig.…”

Section: Resultsmentioning

confidence: 99%

Interaction of structure-specific and promiscuous G-protein–coupled receptors mediates small-molecule signaling in Caenorhabditis elegans

Park

O’Doherty

Somvanshi

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

113

126

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A chemically diverse family of small-molecule signals, the ascarosides, control developmental diapause (dauer), olfactory learning, and social behaviors of the nematode model organism, Caenorhabditis elegans. The ascarosides act upstream of conserved signaling pathways, including the insulin, TGF-β, serotonin, and guanylyl cyclase pathways; however, the sensory processes underlying ascaroside function are poorly understood. Because ascarosides often are multifunctional and show strongly synergistic effects, characterization of their receptors will be essential for understanding ascaroside biology and may provide insight into molecular mechanisms that produce synergistic outcomes in smallmolecule sensing. Based on DAF-8 immunoprecipitation, we here identify two G-protein-coupled receptors, DAF-37 and DAF-38, which cooperatively mediate ascaroside perception. daf-37 mutants are defective in all responses to ascr#2, one of the most potent dauer-inducing ascarosides, although this mutant responds normally to other ascarosides. In contrast, daf-38 mutants are partially defective in responses to several different ascarosides. Through cell-specific overexpression, we show that DAF-37 regulates dauer when expressed in ASI neurons and adult behavior when expressed in ASK neurons. Using a photoaffinity-labeled ascr#2 probe and amplified luminescence assays (AlphaScreen), we demonstrate that ascr#2 binds to DAF-37. Photobleaching fluorescent energy transfer assays revealed that DAF-37 and DAF-38 form heterodimers, and we show that heterodimerization strongly increases cAMP inhibition in response to ascr#2. These results suggest that that the ascarosides' intricate signaling properties result in part from the interaction of highly structure-specific G-protein-coupled receptors such as DAF-37 with more promiscuous G-protein-coupled receptors such as DAF-38. chemosensation | photoaffinity labeling | signal transduction | small-molecule receptor | pheromone signaling

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“…For those inhibitors where the binding site is less understood or completely unknown, new techniques are emerging such as designing cross-linkable derivatives of the antibiotics to make responsible subunits detectable (Bender et al, 2007;Biasotti et al, 2003;Mayer and Maier, 2007;Shen et al, 2005). But as long as a high resolution structure of the complete V-ATPase or at least the V O complex is not available, a major part of this interaction will remain enigmatic.…”

Section: Perspectivesmentioning

confidence: 99%

Inhibitors of V-ATPases: old and new players

Huss

Wieczorek

2009

Journal of Experimental Biology

193

189

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SummaryV-ATPases constitute a ubiquitous family of heteromultimeric, proton translocating proteins. According to their localization in a multitude of eukaryotic endomembranes and plasma membranes, they energize many different transport processes. Currently, a handful of specific inhibitors of the V-ATPase are known, which represent valuable tools for the characterization of transport processes on the level of tissues, single cells or even purified proteins. The understanding of how these inhibitors function may provide a basis to develop new drugs for the benefit of patients suffering from diseases such as osteoporosis or cancer. For this purpose, it appears absolutely essential to determine the exact inhibitor binding site in a target protein on the one side and to uncover the crucial structural elements of an inhibitor on the other side. However, even for some of the most popular and long known V-ATPase inhibitors, such as bafilomycin or concanamycin, the authentic structures of their binding sites are elusive. The aim of this review is to summarize the recent advances for the old players in the inhibition game, the plecomacrolides bafilomycin and concanamycin, and to introduce some of the new players, the macrolacton archazolid, the benzolactone enamides salicylihalamide, lobatamide, apicularen, oximidine and cruentaren, and the indolyls.

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Design and Synthesis of a Tag‐Free Chemical Probe for Photoaffinity Labeling

Cited by 50 publications

References 59 publications

Synthesis and properties of oligonucleotides that contain a triazole‐linked nucleic acid dimer

Synthesis and properties of oligonucleotides that contain a triazole‐linked nucleic acid dimer

Interaction of structure-specific and promiscuous G-protein–coupled receptors mediates small-molecule signaling in Caenorhabditis elegans

Inhibitors of V-ATPases: old and new players

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