title: Fluorescent mimetics of CMP-Neu5Ac are highly potent, cell-permeable polarization probes of eukaryotic and bacterial sialyltransferases and inhibit cellular sialylation.
abstract: Oligosaccharides of the glycolipids and glycoproteins at the outer membranes of human cells carry terminal neuraminic acids, which are responsible for recognition events and adhesion of cells, bacteria, and virus particles. The synthesis of neuraminic acid containing glycosides is accomplished by intracellular sialyl transferases. Therefore, the chemical manipulation of cellular sialylation could be very important to interfere with cancer development, inflammations, and infections. The development and applications of the first nanomolar fluorescent inhibitors of sialyl transferases are described herein. The obtained carbohydrate-nucleotide mimetics were found to bind all four commercially available and tested eukaryotic and bacterial sialyl transferases in a fluorescence polarization assay. Moreover, it was observed that the anionic mimetics intruded rapidly and efficiently into cells in vesicles and translocated to cellular organelles surrounding the nucleus of CHO cells. The new compounds inhibit cellular sialylation in two cell lines and open new perspectives for investigations of cellular sialylation.
authors.affiliation: Medicinal Chemistry, Freie Universität Berlin, Königin-Luise-Strasse 2+4, 14195 Berlin (Germany) http://www.bcp.fu-berlin.de/ag-rademann; Department of Medicinal Chemistry, Leibniz Institut für Molekulare Pharmakologie (FMP), Robert-Rössle-Strasse 10, 13125 Berlin (Germany).
authors.forename: Johannes J
authors.initials: JJ
authors.lastname: Preidl
authors.forename: Vinayaga S
authors.initials: VS
authors.lastname: Gnanapragassam
authors.forename: Michael
authors.initials: M
authors.lastname: Lisurek
authors.forename: Jörn
authors.initials: J
authors.lastname: Saupe
authors.forename: Rüdiger
authors.initials: R
authors.lastname: Horstkorte
authors.forename: Jörg
authors.initials: J
authors.lastname: Rademann
mesh_terms: D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D002463:Cell Membrane Permeability; D006224:Cricetinae; D003412:Cricetulus; D003568:Cytidine Monophosphate / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism*; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D062105:Molecular Docking Simulation; D009438:Neuraminic Acids / Q000737:chemistry / Q000378:metabolism; D016979:Pasteurella multocida / Q000201:enzymology; D010776:Photobacterium / Q000201:enzymology; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012794:Sialic Acids / Q000737:chemistry* / Q000378:metabolism; D012799:Sialyltransferases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D009438:Neuraminic Acids; D012794:Sialic Acids; C561601:cytidine-5'-monophosphosialic acid; D012799:Sialyltransferases; D003568:Cytidine Monophosphate
keywords: fluorescence polarization; metastasis; neuraminic acid; protein-binding probes; sialyltransferases