On-cell saturation transfer difference NMR for the identification of FimH ligands and inhibitors

Published on Jul 1, 2021in Bioorganic Chemistry4.831
· DOI :10.1016/J.BIOORG.2021.104876
Alessandro Palmioli14
Estimated H-index: 14
Paola Sperandeo18
Estimated H-index: 18
+ 2 AuthorsCristina Airoldi26
Estimated H-index: 26
Abstract We describe the development of an on-cell NMR method for the rapid screening of FimH ligands and the structural identification of ligand binding epitopes. FimH is a mannose-binding bacterial adhesin expressed at the apical end of type 1 pili of uropathogenic bacterial strains and responsible for their d -mannose sensitive adhesion to host mammalian epithelial cells. Because of these properties, FimH is a key virulence factor and an attractive therapeutic target for urinary tract infection. We prepared synthetic d -mannose decorated dendrimers, we tested their ability to prevent the FimH-mediated yeast agglutination, and thus we used the compounds showing the best inhibitory activity as models of FimH multivalent ligands to set up our NMR methodology. Our experimental protocol, based on on-cell STD NMR techniques, is a suitable tool for the screening and the epitope mapping of FimH ligands aimed at the development of new antiadhesive and diagnostic tools against urinary tract infection pathogens. Notably, the study is carried out in a physiological environment, i.e. at the surface of living pathogen cells expressing FimH.
#1Natasha E. Hatton (Ebor: University of York)H-Index: 1
#2Christoph G. Baumann (Ebor: University of York)H-Index: 21
Last. Martin A. Fascione (Ebor: University of York)H-Index: 13
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During their lifetime almost half of women will experience a symptomatic urinary tract infection (UTI) with a further half experiencing a relapse within six months. Currently UTIs are treated with antibiotics, but increasing antibiotic resistance rates highlight the need for new treatments. Uropathogenic Escherichia coli (UPEC) is responsible for the majority of symptomatic UTI cases and thus has become a key pathological target. Adhesion of type one pilus subunit FimH at the surface of UPEC str...
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#1Jenny Hall (UEA: University of East Anglia)H-Index: 2
#2Azmat Sohail (Medical University of Vienna)H-Index: 3
Last. Fraser MacMillan (UEA: University of East Anglia)H-Index: 28
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Saturation-transfer difference (STD) NMR spectroscopy is a fast and versatile method which can be applied for drug-screening purposes, allowing the determination of essential ligand binding affinities (KD). Although widely employed to study soluble proteins, its use remains negligible for membrane proteins. Here the use of STD NMR for KD determination is demonstrated for two competing substrates with very different binding affinities (low nanomolar to millimolar) for an integral membrane transpo...
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#2Payam Behzadi (IAU: Islamic Azad University)H-Index: 13
Last. Daniela Scribano (Sapienza University of Rome)H-Index: 12
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Chaperone-usher fimbrial adhesins are powerful weapons against the uropathogens that allow the establishment of urinary tract infections (UTIs). As the antibiotic therapeutic strategy has become less effective in the treatment of uropathogen-related UTIs, the anti-adhesive molecules active against fimbrial adhesins, key determinants of urovirulence, are attractive alternatives. The best-characterized bacterial adhesin is FimH, produced by uropathogenic Escherichia coli (UPEC). Hence, a number of...
13 CitationsSource
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#2Cecilia Ceresa (University of Milano-Bicocca)H-Index: 14
Last. Cristina Airoldi (University of Milano-Bicocca)H-Index: 26
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Abstract We report the NMR characterization of the molecular interaction between Gastrin Releasing Peptide Receptor (GRP-R) and its natural ligand bombesin (BN). GRP-R is a transmembrane G-protein coupled receptor promoting the stimulation of cancer cell proliferation; in addition, being overexpressed on the surface of different human cancer cell lines, it is ideal for the development of new strategies for the selective targeted delivery of anticancer drugs and diagnostic devices to tumor cells....
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#1Alessandro Palmioli (University of Milano-Bicocca)H-Index: 14
#2Paola Sperandeo (University of Milan)H-Index: 18
Last. Cristina Airoldi (University of Milano-Bicocca)H-Index: 26
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#1Jana Mrázková (Masaryk University)H-Index: 2
#2Lenka Malinovská (Masaryk University)H-Index: 6
Last. Michaela Wimmerová (Masaryk University)H-Index: 31
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: Lectins are a group of ubiquitous proteins which specifically recognize and reversibly bind sugar moieties of glycoprotein and glycolipid constituents on cell surfaces. The mutagenesis approach is often employed to characterize lectin binding properties. As lectins are not enzymes, it is not easy to perform a rapid specificity screening of mutants using chromogenic substrates. It is necessary to use different binding assays such as isothermal titration calorimetry (ITC), surface plasmon resona...
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#1Alessandro Palmioli (University of Milano-Bicocca)H-Index: 14
#2Sara Bertuzzi (University of Milano-Bicocca)H-Index: 2
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Abstract The growing interest in medicinal plants for the identification of new bioactive compounds and the formulation of new nutraceuticals and drugs prompted us to develop a powerful experimental approach allowing the detailed metabolic profiling of complex plant extracts, the identification of ligands of macromolecular targets of biomedical relevance and a preliminary characterization of their biological activity. To this end, we selected Peucedanum ostruthium, a plant traditionally employed...
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The design, synthesis and characterization of novel glycoconjugate luminescent probes based on dinuclear rhenium(I) complexes are reported. A multivalent platform bearing different carbohydrate moieties (Glc, Gal and Man) was used to target carbohydrate-binding proteins (lectins), exploiting the unique photophysical characteristics of a Re(I) luminophore for protein sensing. Our results show that these glycoconjugates, non-luminescent in aqueous medium, are able to specifically bind different le...
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An efficient study of carbohydrate-protein interactions was achieved using multivalent glycodendrimer library. Different dendrimers with varied peripheral sugar densities and linkers provided an arsenal of potential novel therapeutic agents that could be useful for better specific action and greater binding affinities against their cognate protein receptors. Highly effective click chemistry represents the basic method used for the synthesis of mannosylated dendrimers. To this end, we used propar...
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