title: Enzymatic synthesis of dimeric glycomimetic ligands of NK cell activation receptors.
abstract: This work reveals new structural relationships in the complex process of the interaction between activation receptors of natural killer cells (rat NKR-P1, human CD69) and novel bivalent carbohydrate glycomimetics. The length, glycosylation pattern and linker structure of receptor ligands were examined with respect to their ability to precipitate the receptor protein from solution, which simulates the in vivo process of receptor aggregation during NK cell activation. It was found that di-LacdiNAc triazole compounds show optimal performance, reaching up to 100% precipitation of the present protein receptors, and achieving high immunostimulatory activities without any tendency to trigger activation-induced apoptosis. In the synthesis of the compounds tested, two enzymatic approaches were applied. Whereas a β-N-acetylhexosaminidase could only glycosylate one of the two acceptor sites available with yields below 10%, the Y284L mutant of human placental β1,4-galactosyltransferase-1 worked as a perfect synthetic tool, accomplishing even quantitative glycosylation at both acceptor sites and with absolute regioselectivity for the C-4 position. This work insinuates new directions for further ligand structure optimisation and demonstrates the strong synthetic potential of the mutant human placental β1,4-galactosyltransferase-1 in the synthesis of multivalent glycomimetics and glycomaterials.
authors.affiliation: Institute of Microbiology, Academy of Sciences of Czech Republic, Vídeňská 1083, CZ-14220 Praha 4, Czech Republic.
authors.forename: Anna
authors.initials: A
authors.lastname: Drozdová
authors.forename: Pavla
authors.initials: P
authors.lastname: Bojarová
authors.forename: Karel
authors.initials: K
authors.lastname: Křenek
authors.forename: Lenka
authors.initials: L
authors.lastname: Weignerová
authors.forename: Birgit
authors.initials: B
authors.lastname: Henssen
authors.forename: Lothar
authors.initials: L
authors.lastname: Elling
authors.forename: Helle
authors.initials: H
authors.lastname: Christensen
authors.forename: Henrik H
authors.initials: HH
authors.lastname: Jensen
authors.forename: Helena
authors.initials: H
authors.lastname: Pelantová
authors.forename: Marek
authors.initials: M
authors.lastname: Kuzma
authors.forename: Karel
authors.initials: K
authors.lastname: Bezouška
authors.forename: Monika
authors.initials: M
authors.lastname: Krupová
authors.forename: David
authors.initials: D
authors.lastname: Adámek
authors.forename: Kristýna
authors.initials: K
authors.lastname: Slámová
authors.forename: Vladimír
authors.initials: V
authors.lastname: Křen
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000276:immunology / Q000378:metabolism; D000945:Antigens, Differentiation, T-Lymphocyte / Q000276:immunology / Q000378:metabolism; D001665:Binding Sites / Q000187:drug effects / Q000276:immunology; D032701:Biomimetics / Q000379:methods*; D005260:Female; D005700:Galactosyltransferases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D047468:Immunoprecipitation; D007694:Killer Cells, Natural / Q000737:chemistry / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D037181:Lectins, C-Type / Q000819:agonists / Q000276:immunology / Q000378:metabolism; D008024:Ligands; D008213:Lymphocyte Activation / Q000187:drug effects / Q000276:immunology; D018716:Molecular Mimicry; D009154:Mutation; D010920:Placenta / Q000201:enzymology; D011134:Polysaccharides / Q000138:chemical synthesis / Q000494:pharmacology; D011247:Pregnancy; D011485:Protein Binding / Q000187:drug effects / Q000276:immunology; D051381:Rats; D055607:Receptors, Natural Killer Cell / Q000819:agonists / Q000276:immunology / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism; D000099192:CD69 Antigens
chemical_list: D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D005700:Galactosyltransferases; D037181:Lectins, C-Type; D008024:Ligands; D011134:Polysaccharides; D055607:Receptors, Natural Killer Cell; D011994:Recombinant Proteins; D001619:beta-N-Acetylhexosaminidases; D000099192:CD69 Antigens; C493310:beta1,4-galactosyltransferase, human