Enzymatic synthesis of dimeric glycomimetic ligands of NK cell activation receptors

Drozdová, Anna; Bojarová, Pavla; Křenek, Karel; Weignerová, Lenka; Henßen, Birgit; Elling, Lothar; Christensen, Helle; Kuzma, Marek; Bezouška, Karel; Krupová, Monika; Adámek, David; Slámová, Kristýna; Křen, Vladimı́r

doi:10.1016/j.carres.2011.04.043

Cited by 26 publications

(15 citation statements)

References 42 publications

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“…The respective structures are depicted in Scheme 3 . The use of click chemistry for attaching intact functionalized oligosaccharides to a multivalent carrier is generally much more advantageous, better defined and higher yielding than attempts to enzymatically glycosylate a multivalent precursor, which often afford a partially glycosylated by-product [ 47 ]. The molecular weights of the prepared glyconanomaterials increased when compared to the respective polymer precursors as a consequence of the presence of the saccharide moieties attached to the polymer structure (cf.…”

Section: Resultsmentioning

confidence: 99%

Biocompatible glyconanomaterials based on HPMA-copolymer for specific targeting of galectin-3

et al. 2018

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BackgroundGalectin-3 (Gal-3) is a promising target in cancer therapy with a high therapeutic potential due to its abundant localization within the tumor tissue and its involvement in tumor development and proliferation. Potential clinical application of Gal-3-targeted inhibitors is often complicated by their insufficient selectivity or low biocompatibility. Nanomaterials based on N-(2-hydroxypropyl)methacrylamide (HPMA) nanocarrier are attractive for in vivo application due to their good water solubility and lack of toxicity and immunogenicity. Their conjugation with tailored carbohydrate ligands can yield specific glyconanomaterials applicable for targeting biomedicinally relevant lectins like Gal-3.ResultsIn the present study we describe the synthesis and the structure-affinity relationship study of novel Gal-3-targeted glyconanomaterials, based on hydrophilic HPMA nanocarriers. HPMA nanocarriers decorated with varying amounts of Gal-3 specific epitope GalNAcβ1,4GlcNAc (LacdiNAc) were analyzed in a competitive ELISA-type assay and their binding kinetics was described by surface plasmon resonance. We showed the impact of various linker types and epitope distribution on the binding affinity to Gal-3. The synthesis of specific functionalized LacdiNAc epitopes was accomplished under the catalysis by mutant β-N-acetylhexosaminidases. The glycans were conjugated to statistic HPMA copolymer precursors through diverse linkers in a defined pattern and density using Cu(I)-catalyzed azide–alkyne cycloaddition. The resulting water-soluble and structurally flexible synthetic glyconanomaterials exhibited affinity to Gal-3 in low μM range.ConclusionsThe results of this study reveal the relation between the linker structure, glycan distribution and the affinity of the glycopolymer nanomaterial to Gal-3. They pave the way to specific biomedicinal glyconanomaterials that target Gal-3 as a therapeutic goal in cancerogenesis and other disorders.Electronic supplementary materialThe online version of this article (10.1186/s12951-018-0399-1) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Biocompatible glyconanomaterials based on HPMA-copolymer for specific targeting of galectin-3

et al. 2018

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“…NMR titrations of mannose, p NP-GlcNAc, 1,2- bis [ N -(2-acetamido-2-deoxy-β- d -glucopyranosyl)- thioureido]-decane [26] and the LELTE peptide with NKR-P1A and CD69 were performed at 25 °C in molar ratios of 1:1 and 1:2 (protein:ligand). In all cases the 1 H- 15 N HSQC spectra were measured and compared with the 1 H- 15 N HSQC spectrum of the protein without the ligand.…”

Section: Resultsmentioning

confidence: 99%

“…Both receptors appeared to have extraordinary affinity for certain disaccharides such as ManNAc(1→4)Glc [20] or deoxynorijimycin and its hexosaminyl derivates [21] as well as negatively charged oligosaccharides and glycosides [22–24]. Monovalent and bivalent LacdiNAc glycomimetics prepared via enzymatic synthesis were also found to be efficient NK cell activators [25,26]. In addition, carboxylated calixarenes [27], glycosyl 1,2,3-triazoles [28] and glycoconjugates of the LELTE peptide [29] have been reported as new prominent classes of high-affinity ligands.…”

Section: Introductionmentioning

confidence: 99%

Re-Evaluation of Binding Properties of Recombinant Lymphocyte Receptors NKR-P1A and CD69 to Chemically Synthesized Glycans and Peptides

Rozbeský

Krejzová

Křenek

et al. 2014

IJMS

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The binding of monosaccharides and short peptides to lymphocyte receptors (human CD69 and rat NKR-P1A) was first reported in 1994 and then in a number of subsequent publications. Based on this observation, numerous potentially high-affinity saccharide ligands have been synthesized over the last two decades in order to utilize their potential in antitumor therapy. Due to significant inconsistencies in their reported binding properties, we decided to re-examine the interaction between multiple ligands and CD69 or NKR-P1A. Using NMR titration and isothermal titration calorimetry we were unable to detect the binding of the tested ligands such as N-acetyl-d-hexosamines and oligopeptides to both receptors, which contradicts the previous observations published in more than twenty papers over the last fifteen years.

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“…A few examples of successful reactions with A:D ratios <1 (as low as 1:10) have been reported (Tables 4, 5 and 8) [20,24,61,85,88,100,[118][119][120] using carbohydrates as acceptors. The most extreme A:D ratio of 1:5000 was used in a thioglycoligase reaction carried out with a mutant of the hexosaminidase from Streptomyces plicatus, which was able to S-glycosylate small peptides and proteins [24].…”

Section: Acceptor:donor Ratiomentioning

confidence: 99%

β-N-Acetylhexosaminidases for Carbohydrate Synthesis via Trans-Glycosylation

et al. 2020

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β-N-acetylhexosaminidases (EC 3.2.1.52) are retaining hydrolases of glycoside hydrolase family 20 (GH20). These enzymes catalyze hydrolysis of terminal, non-reducing N-acetylhexosamine residues, notably N-acetylglucosamine or N-acetylgalactosamine, in N-acetyl-β-D-hexosaminides. In nature, bacterial β-N-acetylhexosaminidases are mainly involved in cell wall peptidoglycan synthesis, analogously, fungal β-N-acetylhexosaminidases act on cell wall chitin. The enzymes work via a distinct substrate-assisted mechanism that utilizes the 2-acetamido group as nucleophile. Curiously, the β-N-acetylhexosaminidases possess an inherent trans-glycosylation ability which is potentially useful for biocatalytic synthesis of functional carbohydrates, including biomimetic synthesis of human milk oligosaccharides and other glycan-functionalized compounds. In this review, we summarize the reaction engineering approaches (donor substrate activation, additives, and reaction conditions) that have proven useful for enhancing trans-glycosylation activity of GH20 β-N-acetylhexosaminidases. We provide comprehensive overviews of reported synthesis reactions with GH20 enzymes, including tables that list the specific enzyme used, donor and acceptor substrates, reaction conditions, and details of the products and yields obtained. We also describe the active site traits and mutations that appear to favor trans-glycosylation activity of GH20 β-N-acetylhexosaminidases. Finally, we discuss novel protein engineering strategies and suggest potential “hotspots” for mutations to promote trans-glycosylation activity in GH20 for efficient synthesis of specific functional carbohydrates and other glyco-engineered products.

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Enzymatic synthesis of dimeric glycomimetic ligands of NK cell activation receptors

Cited by 26 publications

References 42 publications

Biocompatible glyconanomaterials based on HPMA-copolymer for specific targeting of galectin-3

Biocompatible glyconanomaterials based on HPMA-copolymer for specific targeting of galectin-3

Re-Evaluation of Binding Properties of Recombinant Lymphocyte Receptors NKR-P1A and CD69 to Chemically Synthesized Glycans and Peptides

β-N-Acetylhexosaminidases for Carbohydrate Synthesis via Trans-Glycosylation

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