- Rapid glycoconjugation with glycosyl amines
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Conjugation of unprotected carbohydrates to surfaces or probes by chemoselective ligation reactions is indispensable for the elucidation of their numerous biological functions. In particular, the reaction with oxyamines leading to the formation of carbohydrate oximes which are in equilibrium with cyclic N-glycosides (oxyamine ligation) has an enormous impact in the field. Although highly chemoselective, the reaction is rather slow. Here, we report that the oxyamine ligation is significantly accelerated without the need for a catalyst when starting with glycosyl amines. Reaction rates are increased up to 500-fold compared to the reaction of the reducing carbohydrate. For comparison, aniline-catalyzed oxyamine ligation is only increased 3.8-fold under the same conditions. Glycosyl amines from mono- and oligosaccharides are easily accessible from reducing carbohydrates via the corresponding azides by using Shoda's reagent (2-chloro-1,3-dimethylimidazolinium chloride, DMC) and subsequent reduction. Furthermore, glycosyl amines are readily obtained by enzymatic release from N-glycoproteins making the method suited for glycomic analysis of these glycoconjugates which we demonstrate employing RNase B. Oxyamine ligation of glycosyl amines can be carried out at close to neutral conditions which makes the procedure especially valuable for acid-sensitive oligosaccharides. This journal is
- Baudendistel, Oliver R.,Rapp, Mareike A.,Steiner, Ulrich E.,Wittmann, Valentin
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p. 14901 - 14906
(2021/12/02)
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- Total Synthesis of Glycosylated Human Interferon-γ
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Interferon-γ(IFN-γ) is a glycoprotein that is responsible for orchestrating numerous critical immune induction and modulation processes and is used clinically for the treatment of a number of diseases. Herein, we describe the total chemical synthesis of homogeneously glycosylated variants of human IFN-γusing a tandem diselenide-selenoester ligation-deselenization strategy in the C- to N-terminal direction. The synthetic glycoproteins were successfully folded, and the structures and antiviral functions were assessed.
- Ashhurst, Anneliese S.,Dowman, Luke J.,Fairbanks, Antony J.,Kwan, Ann,Larance, Mark,Li, Henry Y.,Payne, Richard J.,Wang, Xiaoyi,Watson, Emma E.
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supporting information
p. 6863 - 6867
(2020/09/15)
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- A Tripeptide Approach to the Solid-Phase Synthesis of Peptide Thioacids and N-Glycopeptides
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A general and robust method for the incorporation of aspartates with a thioacid side chain into peptides has been developed. Pseudoproline tripeptides served as building blocks for the efficient fluorenylmethyloxycarbonyl (Fmoc) solid-phase synthesis of t
- Sch?we, Markus Julian,Keiper, Odin,Unverzagt, Carlo,Wittmann, Valentin
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p. 15759 - 15764
(2019/11/19)
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- N-Linked Glycosyl Auxiliary-Mediated Native Chemical Ligation on Aspartic Acid: Application towards N-Glycopeptide Synthesis
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A practical approach towards N-glycopeptide synthesis using an auxiliary-mediated dual native chemical ligation (NCL) has been developed. The first NCL connects an N-linked glycosyl auxiliary to the thioester side chain of an N-terminal aspartate oligopeptide. This intermediate undergoes a second NCL with a C-terminal thioester oligopeptide. Mild cleavage provides the desired N-glycopeptide.
- Chai, Hua,Le Mai Hoang, Kim,Vu, Minh Duy,Pasunooti, Kalyan,Liu, Chuan-Fa,Liu, Xue-Wei
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supporting information
p. 10363 - 10367
(2016/08/24)
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- Convergent Synthesis of N-Linked Glycopeptides via Aminolysis of ω-Asp p-Nitrophenyl Thioesters in Solution
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An efficient N-linked glycosylation reaction between glycosylamines and p-nitrophenyl thioester peptides has been developed. The reaction conditions are mild and compatible with the C-terminal free carboxylic acid group and the unprotected N-linked sialyl
- Du, Jing-Jing,Gao, Xiao-Fei,Xin, Ling-Ming,Lei, Ze,Liu, Zheng,Guo, Jun
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supporting information
p. 4828 - 4831
(2016/10/14)
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- Rapid formation of N-glycopeptides via Cu(II)-promoted glycosylative ligation
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Herein is described the chemoselective Cu(II)-HOBt promoted chemical ligation of glycosylamines and peptide thioacids to give N-glycosylated peptides. The method is distinguished from other chemical approaches to peptide N-glycosylation in that (1) it can be employed in the presence of unprotected N-terminal and Lys side chain amines; (2) it is remarkably fast, going to completion in under 30 min; and (3) it produces glycopeptides without attendant aspartimide formation.
- Joseph, Ryan,Dyer, Frank Brock,Garner, Philip
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p. 732 - 735
(2013/03/29)
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- COMPOSITIONS, METHODS OF SYNTHESIS AND USE OF CARBOHYDRATE TARGETED AGENTS
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The invention provides D02S derivatives conjugated to monosaccharide ligands directly or through a linker and optionally chelated to a metal, wherein theD02S derivatives having the following structure: wherein R1', R2' are each independently-OH or -O-alkyl; R1 is a hydrogen, a linker, or a ligand; R3 is a linker and/or a ligand; and n is an integer from 1 to 10; the linker is an amino acid, a peptide, an amino alcohol, a polyethylylene glycol, an alkyl, an alkenyl, an alkynyl, an azide, an aromatic compound, a carboxylic acid, or an ester, the alkyl, alkenyl, or alkynyl is optionally substituted with an alkyl, a halogen, a nitro group, a hydroxyl group, an amino group, or a carboxyl group; the ligand is a GLUT1 targeting moiety.
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Page/Page column 13
(2013/02/28)
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- Mixed SAMs and MALDI-ToF MS: Preparation of N-glycosylamine derivative and thioctic acid methyl ester bearing 1,2-dithiolane groups and detection of enzymatic reaction on Au
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Herein, we report an enzymatic galactosylation reaction of β-glucopyranosylamide 4 and thioctic acid methyl ester 5 bearing 1,2-dithiolane groups to form a new system of mixed self-assembled monolayers (SAMs) on gold. Characterization of the enzymatic activity was conveniently achieved by mass spectrometry.
- Hassan, Hani Mutlak A.,Maltman, Beatrice A.
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experimental part
p. 6 - 9
(2012/03/26)
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- Synthesis of amino-bridged oligosaccharide mimetics
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Synthesis of amino-bridged oligosaccharides using reductive animation opens rapid access to novel glycomimetic target structures as potential ligands for the receptor protein NKR P1 of natural killer cells. Emphasis was laid on fast and facile synthetic routes. The carbonyl building blocks were easily obtained by oxidation with Dess-Martin periodinane or iodoxybenzoic acid (IBX). For the required amino-function-alized units, reduction of azide precursors was advantageous, and generation of the novel oligosaccharides was achieved by subsequent reductive amination. The target saccharide structures feature a bridging nitrogen atom inserted between two non-anomeric positions as well as including one anomeric position.
- Neumann, Janna,Thiem, Joachim
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scheme or table
p. 900 - 908
(2010/04/23)
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- Study of on-resin convergent synthesis of N-linked glycopeptides containing a large high mannose N-linked oligosaccharide
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Here we present a convergent on-resin glycosylamine coupling strategy for solid phase N-linked glycopeptide synthesis, and apply it to the synthesis of high mannose containing glycopeptides. In this strategy, the 2-phenylisopropyl protecting group is used as an orthogonal handle to create glycosylation sites on-resin after synthesis of nonglycosylated peptides. In addition to allowing selective deprotection of aspartic acid residues for creation of glycosylation sites, the 2-phenylisopropyl protecting group also efficiently suppresses aspartimide formation during peptide synthesis. The key step of on-resin glycosylamine coupling to an aspartic acid residue was first optimized for a small sugar, N-acetylglucosamine, and then applied to a much larger high mannose oligosaccharide, Man8GlcNAc2. Satisfying coupling yields were obtained for both small and large sugars. The use of on-resin glycosylamine coupling simplifies purification of N-linked glycopeptides, and also allows convenient recovery of unreacted valuable large oligosaccharides. This approach was applied to the solid phase synthesis of glycosylated forms of the 34 amino acid HIV-1 gp41 C34 glycopeptide, which is an HIV-1 entry inhibitor. The HIV-1 entry inhibition assay of synthesized glycopeptides showed the retention of bioactivity of high mannose Man8GlcNAc2-C34.
- Chen, Rui,Tolbert, Thomas J.
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experimental part
p. 3211 - 3216
(2010/05/14)
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- Efficient use of the Dmab protecting group: Applications for the solid-phase synthesis of N-linked glycopeptides
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An efficient protocol for the chemoselective removal of Dmab esters on the solid phase is reported; this method has been successfully utilised for the convergent solid phase synthesis of N-linked glycopeptides. The Royal Society of Chemistry 2009.
- Conroy, Trent,Jolliffe, Katrina A.,Payne, Richard J.
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supporting information; experimental part
p. 2255 - 2258
(2009/09/26)
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- Chemoenzymatic synthesis and lectin array characterization of a class of N-glycan clusters
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N-Glycans are major components of many glycoproteins. These sugar moieties are frequently involved in important physiological and disease processes via their interactions with a variety of glycanbinding proteins (GBP). Clustering effect is an important feature in many glycan-lectin interactions. We describe in this paper a chemoenzymatic synthesis of novel N-glycan clusters using a tandem endoglycosidase-catalyzed transglycosylation. It was found that the internal β-1,2-linked GlcNAc moieties in the N-glycan core, once exposed in the nonreducing terminus, was able to serve as acceptors for transglycosylation catalyzed by Endo-A and EndoM-N175A. This efficient chemoenzymatic method allows a quick extension of the sugar chains to form a class of glycan clusters in which sugar residues are all connected by native glycosidic linkages found in natural N-glycans. In addition, a discriminative enzymatic reaction at the two GlcNAc residues could be fulfilled to afford novel hybrid clusters. Lectin microarray studies revealed unusual properties in glyco-epitope expression by this panel of structurally well-defined synthetic N-glycans. These new compounds are likely valuable for functional glycomics studies to unveil new functions of both glycans and carbohydrate-binding proteins.
- Huang, Wei,Wang, Denong,Yamada, Masao,Wang, Lai-Xi
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experimental part
p. 17963 - 17971
(2010/03/30)
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- Synthesis of neoglycoconjugates by the desulfurative rearrangement of allylic disulfides
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(Chemical Equation Presented) Two series of neoglycosyl donors are prepared on the basis of connection of an allylic disulfide motif to the anomeric center via a simple O-glycosyl linkage or N-glycosyl amide unit. Conjugation of both sets of donors to cysteine in peptides is demonstrated through classical disulfide exchange followed by the phosphine-mediated desulfurative allylic rearrangement resulting in neoglycopeptides characterized by a simple thioether spacer. The conjugation reaction functions in the absence of protecting groups on both the neoglycosyl donor and peptide in aqueous media at room temperature.
- Crich, David,Yang, Fan
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experimental part
p. 7017 - 7027
(2009/05/09)
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- α-Glucopyranoimidazolines as intermediate analogue inhibitors of family 20 β-N-acetylglucosaminidases
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The α-glucopyranoimidazolines, 2-methyl-(1,2-dideoxy-α-D- glucopyrano)[2,1-d]-1-imidazolines 1 and 2, have been synthesized and evaluated as inhibitors of β-N-acetylglucosaminidases (NAGs). Compounds 1 and 2, mimicking the oxazolinium ion intermediate in
- Kato, Masahiro,Uno, Tetsuya,Hiratake, Jun,Sakata, Kanzo
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p. 1563 - 1571
(2007/10/03)
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- An efficient synthetic route to glycoamino acid building blocks for glycopeptide synthesis
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(Chemical Equation Presented) Chemical glycopeptide synthesis requires access to gram quantities of glycosylated amino acid building blocks. Hence, the efficiency of synthesis of such building blocks is of great importance. Here, we report a fast and high
- Bejugam, Mallesham,Flitsch, Sabine L.
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p. 4001 - 4004
(2007/10/03)
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- Tight binding ligand approach to oligosaccharide-grafted protein
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A novel type of artificial glycoprotein was developed, by using dihydrofolate reductase (DHFR) and methotrexate (MTX) as a protein-ligand pair. Various oligosaccharides linked to MTX were shown to bind tightly with DHFR and afforded oligosaccharide-grafted protein, which could be isolated easily by lectin beads.
- Totani, Kiichiro,Matsuo, Ichiro,Ito, Yukishige
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p. 2285 - 2289
(2007/10/03)
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- An expeditious route to GlcNAc-Cbz-Asn by chemo-enzymatic synthesis
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A short-step route to GlcNAc-Cbz-Asn was developed. Treatment of GlcNAc in sat. aq. NH4HCO3 solution and subsequent electorodialytic desalting provided ammonia-free glycosylamine in large quantity. The product was coupled with Cbz-As
- Tokuda,Takahashi,Matoishi,Ito,Sugai
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- Chemoselective ligation of maleimidosugars to peptides/protein for the preparation of neoglycopeptides/neoglycoprotein
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Two types of maleimidosugars as thiol-selective carbohydrates, 1-maleimidosugars and acetyl-linked maleimidosugars, were efficiently synthesized. They were coupled to glutathione, Fas peptide and bovine serium albumin (BSA) to prepare the corresponding glycosylated peptides and a protein via stable thioether linkages in a chemoselective manner.
- Shin, Injae,Jung, Hyuk-Jun,Lee, Myung-Ryul
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p. 1325 - 1328
(2007/10/03)
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- Synthesis of N4-(2-acetamido-2-deoxy-β-D-glucopyranosyl)-L-asparagine analogues. Complete NMR assignments of chloroacetamide, bromoacetamide and glycinamide analogues
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N4-(2-Acetamido-2-deoxy-β-D-glucopyranosyl)-L-asparagine is the principle linkage in the structure of N-linked glycoproteins. Complete assignments of the 1H and 13C NMR spectra for three analogues of the naturally-occurring N-glycosylic structure were made for N-(2-acetamido-2-deoxy-β-D-glucopyranosyl)chloroacetamide, N-(2-acetamido-2-deoxy-β-D-glucopyranosyl)bromoacetamide and N1-(2-acetamido-2-deoxy-β-D-glucopyranosyl)glycinamide. The methylene groups adjacent to the N-glycosylic bond are AB spin systems. Copyright
- Malik, Jayshri J.,Risley, John M.
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- Synthesis of N4-(2-acetamido-2-deoxy-β-D-glucopyranosyl)-L-asparagine analogues. n-Butyramide, 3-chloropropionamide, 3-aminopropionamide, and isovaleramide analogues
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The syntheses of four analogues of N4-(2-acetamido-2-deoxy-β-D-glucopyranosyl)-L-asparagine are described. Activated carboxylic acids were reacted with 2-acetamido-2-deoxy-β-D-glucopyranosylamine. n-Butyric anhydride gave N-(2-acetamido-2-deoxy-β-D-glucopyranosyl)-n-butyramide. 3-Chloropropionic anhydride was synthesized from 3-chloropropionic acid and gave N-(2-acetamido-2-deoxy-β-D-glucopyranosyl)-3-chloropropionamide. Equilibration of the latter with ammonium bicarbonate gave N1-(2-acetamido-2-deoxy-β-D-glucopyranosyl)-3- aminopropionamide. Succinimidyl isovalerate was synthesized and gave N-(2-acetamido-2-deoxy-β-D-glucopyranosyl)isovaleramide.
- Kaylor, Jerry J.,Risley, John M.
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p. 439 - 444
(2007/10/03)
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- Synthesis of N4-(2-acetamido-2-deoxy-β-D-glucopyranosyl)-L-asparagine analogues. L-2-chloro-, L-2-bromo-, and D,L-2-methylsuccinamic acid analogues
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L-Chlorosuccinic anhydride, L-bromosuccinic anhydride, and D,L-methylsuccinic anhydride react with 2-acetamido-2-deoxy-β-D-glucopyranosylamine to give varying mixtures of N4-(β-GlcNAc)-2-substituted- and N4-(β-GlcNAc)-3-substituted-succinamic acid isomers. The two regioisomers are separated by anion exchange chromatography. The N4-(β-GlcNAc)-2-substituted-succinamic acid isomers are characterized as analogues of N4-(2-acetamido-2-deoxy-β-D-glucopyranosyl)-L-asparagine.
- Xia, Yuan-Qing,Risley, John M.
-
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- Unprotected oligosaccharides as phase tags: solution-phase synthesis of glycopeptides with solid-phase workups.
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[reaction--see text] N-Linked glycopeptides were synthesized from glycosyl asparagines containing unprotected oligosaccharides and other simple amino acids by an Fmoc method. The free oligosaccharide chains were used as phase tags to facilitate the produc
- Wen,Guo
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p. 3773 - 3776
(2007/10/03)
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- Synthesis of N4-(2-acetamido-2-deoxy-β-D-glucopyranosyl)-L-asparagine analogues: Succinamide, L-2-hydroxysuccinamide, and L-2-hydroxysuccinamic acid hydrazide analogues
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The syntheses of three analogues of N4-(2-acetamido-2-deoxy-β-D-glucopyranosyl)-L-asparagine are described. N-(2-Acetamido-2-deoxy-β-D-glucopyranosyl)succinamide was synthesized by the reaction of pentafluorophenyl succinamate with 2-acetamido-
- Huang, De Hua,Risley, John M.
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p. 487 - 493
(2007/10/03)
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- A fluorometric assay for glycosyltransferase activities using sugars aminated and tagged with 7-hydroxycoumarin-3-carboxylic acid as substrates and high performance liquid chromatography
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We developed a novel fluorometric assay method for the measurement of glycosyltransferase activities using mono- and di-saccharides aminated and tagged with 7-hydroxycoumarin-3-carboxylic acid (coumarin) as substrates, N- acetylglucosamine (GlcNAc)-coumarin for β1,4-galactosyltransferase from bovine milk and Galβ1-4GlcNAc-coumarin for α2,3- and α2,6- sialyltransferases from rat liver. Using Galβ1-3GlcNAc and Galβ1-4GlcNAc- coumarin, α1, 3/4 /4- and 1/4 ,3-fucosyltransferase activities were also determined, respectively. These enzymatic products liberated by the reactions of glycosyltransferases in the presence of sugar nucleotides, were separated by a normal phase or an ion-pair reversed phase HPLC with an isocratic elution and fluorescence detection. We applied this assay method to determine the glycosyltransferase activities in 8 kinds of human tumor cell lines, including the cell lines derived from hepatocytes (HuH-7, HepG2), colonic cells (Colo205, HT-29), myelocytes (HL-60, U-937), B-lymphocytes (Daudi) and T-lymphocytes (Jurkat). This assay method is accurate and easy compared with other isotopic and non-isotopic assay methods, and is sensitive enough to measure glycosyltransferase activities in cell homogenates.
- Higai, Koji,Masuda, Daisuke,Matsuzawa, Yukinari,Satoh, Tamae,Matsumoto, Kojiro
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p. 333 - 338
(2007/10/03)
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- Novel sequential solid-phase synthesis of N-linked glycopeptides from natural sources
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In the present report a practical and versatile procedure for the solid-phase synthesis of N-linked glycopeptides from natural sources has been demonstrated. The approach is based on the mild hydrazinolysis procedure to release N-linked oligosaccharides in their intact unreduced form from the natural glycoproteins, e.g. fetuin and ribonuclease B and subsequent formation of the corresponding glycosylamines. Treatment of the reducing sugars 1-7 with a saturated solution of ammonium hydrogen carbonate in either water or dimethyl sulfoxide (DMSO) gives in almost quantitative yields the glycosylamines 8-14. Coupling of the unprotected glycosylamines 8-14 to the side-chain-activated aspartic acid derivative Fmoc-Asp(ODhbt)-OBut 16 affords the N-glycosylated asparagine derivatives 17-23. Subsequent acetylation of the carbohydrate hydroxy groups and cleavage of the tert-Bu ester by trifluoroacetic acid (TFA) treatment yields the glycosylated N-linked building blocks 31-37. The building blocks 31-37 are then incorporated into the multiple-column peptide-synthesis protocol of the glycopeptide T-cell epitope analogues 40-46 of the mouse haemoglobin-derived decapeptide Hb (67-76), VITAFNEGLK. The decapeptide sequence VITAFNEGLK binds well to the MHC Class II Ek molecule and is non-immunogenic in CBA/J mice. Syntheses of several natural and unnatural glycosylations, e.g. N-acetylglucosamine, N,N′-diacetylchitobiose, glucose, maltotriose, maltoheptose and di- and triantennary complex oligosaccharides on the decapeptide Hb (67-76) affording the N-linked glycopeptides 40-46 are described. The N-linked glycopeptides 40-46 have been fully characterised by 1D- and 2D-1H and 13C NMR spectroscopy and by ES-MS.
- Meinjohanns, Ernst,Meldal, Morten,Paulsen, Hans,Dwek, Raymond A.,Bock, Klaus
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p. 549 - 560
(2007/10/03)
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- Synthesis of Artificial N-Glycopolypeptides Carrying N-Acetyllactosamine and Related Compounds and Their Specific Interactions with Lectins
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Artificial N-glycopolypeptides carrying N-acetyllactosamine (LacNAc) or related compounds were synthesized. First, sugars were converted into their corresponding β-glycosylamines with ammonium hydrogen carbonate. Then, the β-glycosylamines were condensated with the carboxyl groups of poly(L-glutamic acid). N-Glycopolypeptides with different degrees of substitution of sugars were isolated by passage through a column of Sephadex G-25. These synthetic polymers were used as model compounds in the analysis of oligosaccharide-lectin interactions. Interactions with some lectins were investigated by agar-gel double-diffusion tests and in terms of inhibition of hemagglutination. A glycopolypeptide substituted with LacNAc reacted with Erythrina cristagalli agglutinin (ECA), peanut (Arachis hypogaea) agglutinin (PNA), Ricinus communis agglutinin-120 (RCA120), wheat germ (Triticum vulgaris) agglutinin (WGA) lectins, which recognize either galactosyl or N-acetylglucosamine (GlcNAc) residues. Other synthetic glycopolymers carrying N-acetylisolactosamine, GlcNAc, N,N′-diacetylchitobiose, or N,N′,N″-triacetylchitotriose also reacted with WGA, and these last two polymers inhibited hemagglutination most. Of these five glycopolypeptides, only the one substituted with LacNAc reacted with ECA. These sugar-substituted glycopolypeptides interacted specifically with the corresponding lectins, no matter how much shorter the sugar side chains of the glycopolymers were than those of natural glycoproteins.
- Zeng, Xiaoxiong,Murata, Takeomi,Kawagishi, Hirokazu,Usui, Taichi,Kobayashi, Kazukiyo
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p. 1171 - 1178
(2007/10/03)
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- Pentafluorophenyl esters for temporary carboxyl group protection in solid phase synthesis of N-linked glycopeptides
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The compound Ac3GlcNAcβ1-NH-Fmoc (3) was synthesized and transformed into the β-glucosyl amine (1) which was subsequently acylated with Fmoc-Asp(Cl)-O-Pfp (5) prepared from the readily available Fmoc-Asp(O-tBu)-O-Pfp (4). The resulting Fmoc-Asn
- Meldal, Morten,Book, Klaus
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p. 6987 - 6990
(2007/12/18)
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- SYNTHESIS OF 2-ACETAMIDO-1-N--2-DEOXY-β-D-GLUCOPYRANOSYLAMINE AND ANALOGS
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2-Acetamido-1-N--2-deoxy-β-D-glucopyranosylamine and analogs containing D-glucopyranosyl, 4-O-β-D-glucopyranosyl-D-glucopyranosyl, L-Phe-L-Ala, and D-Phe-L-Ser were synth
- Tamura, Masahiro,Okai, Hideo
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p. 207 - 218
(2007/10/02)
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