Oxo-ester mediated native chemical ligation on microarrays: an efficient and chemoselective coupling methodology

Weissenborn, Martin J.; Castangia, Roberto; Wehner, Johannes W.; Šardzík, Róbert; Flitsch, Sabine L.

doi:10.1039/c2cc30844d

Cited by 24 publications

(38 citation statements)

References 36 publications

(31 reference statements)

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“…19 Weissenborn et al described OMNCL on oxo-ester activated surfaces and found 2,3,4,5,6-pentafluorophenyl (PFP) to be more efficient than p NP and N -hydroxysuccinimide (NHS) activating agents. 20 …”

Section: Introductionmentioning

confidence: 99%

Hydrogels formed by oxo-ester mediated native chemical ligation

et al. 2013

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Oxo-ester mediated native chemical ligation (OMNCL) is a variation of the more general native chemical ligation (NCL) reaction that is widely employed for chemoselective ligation of peptide fragments. While OMNCL has been used for a variety of peptide ligations and for biomolecular modification of surfaces, it is typically practiced under harsh conditions that are unsuitable for use in a biological context. In this report we describe the use of OMNCL for polymer hydrogel formation, in-vitro cell encapsulation, and in-vivo implantation. Multivalent polymer precursors containing N-hydroxysuccinimide (NHS) activated oxo-esters and N-cysteine (N-Cys) endgroups were chemically synthesized from branched poly(ethylene glycol) (PEG). Hydrogels formed rapidly at physiologic pH upon mixing of aqueous solutions of NHS and N-Cys functionalized PEGs. Quantitative 1H NMR experiments showed that the reaction proceeds through an OMNCL pathway involving thiol capture to form a thioester intermediate, followed by an S-to-N acyl rearrangement to yield an amide cross-link. pH and temperature were found to influence gelation rate, allowing tailoring of gelation times from a few seconds to a few minutes. OMNCL hydrogels initially swelled before contracting to reach an equilibrium increase in relative wet weight of 0%. This unique behavior impacted the gel stiffness and was attributed to latent formation of disulfide cross-links between network-bound Cys residues. OMNCL hydrogels were adhesive to hydrated tissue, generating a lap shear adhesion strength of 46 kPa. Cells encapsulated in OMNCL hydrogels maintained high viability, and in-situ formation of OMNCL hydrogel by subcutaneous injection in mice generated a minimal acute inflammatory response. OMNCL represents a promising strategy for chemical cross-linking of hydrogels in a biological context and is an attractive candidate for in-vivo applications such as wound healing, tissue repair, drug delivery, and tissue engineering.

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Section: Introductionmentioning

confidence: 99%

Hydrogels formed by oxo-ester mediated native chemical ligation

et al. 2013

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“…In addition, mannosidic glycoamino acids are of interest as ligands for mannose-specific lectins in solution [17] as well as for the fabrication of glycoarrays on solid support [10,18–19]. Moreover, we are interested to advance glycoamino acid deriatives such as 3 into ligands that can be “switched” between two states by oxidation/reduction.…”

Section: Resultsmentioning

confidence: 99%

“…This is an attractive concept, because it can be combined with solid-phase peptide synthesis (SPPS) [4–5], as well as with native chemical ligation (NCL) utilizing an S → N acyl shift [3,6–10]. In addition, glycocysteine derivatives can be easily converted into the corresponding dimers by oxidiation of the cysteine moiety into the respective cystine form.…”

Section: Introductionmentioning

confidence: 99%

S-Fluorenylmethyl protection of the cysteine side chain upon N^α-Fmoc deprotection

Wehner

2012

Beilstein J. Org. Chem.

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SummaryDeprotection of an N α-Fmoc-protected glycocysteine derivative 7 with an excess of morpholine unexpectedly led to the fluorenylmethyl-protected thioether 8 in high yield. The suggested mechanism for this reaction comprises the addition of the cysteine thiolate on the exocyclic double bond of dibenzofulvene, which is formed during Fmoc deprotection. The influence of base concentration on this transprotection reaction was studied.

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“…In the case that the E → Z isomerisation process is high-yielding and the lifetime of the ( Z )-form of the azobenzene glycoside is long enough, it can be employed in bacterial adhesion assays independently from the more stable ( E )-isomer. Eventually, this type of azobenzene mannobioside can be further functionalised to be attached to various supports such as oligofunctional core molecules [15] or surfaces, to achieve switchable adhesive surfaces in continuation of our work on glycoarrays [16–18] (Fig. 1).…”

Section: Introductionmentioning

confidence: 99%

Synthesis and testing of the first azobenzene mannobioside as photoswitchable ligand for the bacterial lectin FimH

Chandrasekaran¹,

Kolbe²,

Beiroth³

et al. 2013

Beilstein J. Org. Chem.

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SummaryIn order to allow spatial and temporal control of carbohydrate-specific bacterial adhesion, it has become our goal to synthesise azobenzene mannosides as photoswitchable inhibitors of type 1 fimbriae-mediated adhesion of E. coli. An azobenzene mannobioside 2 was prepared and its photochromic properties were investigated. The E→Z isomerisation was found to be highly effective, yielding a long-lived (Z)-isomer. Both isomers, E and Z, show excellent water solubility and were tested as inhibitors of mannoside-specific bacterial adhesion in solution. Their inhibitory potency was found to be equal and almost two orders of magnitude higher than that of the standard inhibitor methyl mannoside. These findings could be rationalised on the basis of computer-aided docking studies. The properties of the new azobenzene mannobioside have qualified this glycoside to be eventually employed on solid support, in order to fabricate photoswitchable adhesive surfaces.

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Oxo-ester mediated native chemical ligation on microarrays: an efficient and chemoselective coupling methodology

Abstract: We report a highly efficient and selective method for the coupling of peptides and glycoconjugates bearing N-terminal cysteines to activated surfaces. This chemoselective method generates stable amide linkages without using any thiol additives.

Cited by 24 publications

References 36 publications

Hydrogels formed by oxo-ester mediated native chemical ligation

Hydrogels formed by oxo-ester mediated native chemical ligation

S-Fluorenylmethyl protection of the cysteine side chain upon N^α-Fmoc deprotection

Synthesis and testing of the first azobenzene mannobioside as photoswitchable ligand for the bacterial lectin FimH

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Oxo-ester mediated native chemical ligation on microarrays: an efficient and chemoselective coupling methodology

Abstract: We report a highly efficient and selective method for the coupling of peptides and glycoconjugates bearing N-terminal cysteines to activated surfaces. This chemoselective method generates stable amide linkages without using any thiol additives.

Cited by 24 publications

References 36 publications

Hydrogels formed by oxo-ester mediated native chemical ligation

Hydrogels formed by oxo-ester mediated native chemical ligation

S-Fluorenylmethyl protection of the cysteine side chain upon Nα-Fmoc deprotection

Synthesis and testing of the first azobenzene mannobioside as photoswitchable ligand for the bacterial lectin FimH

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S-Fluorenylmethyl protection of the cysteine side chain upon N^α-Fmoc deprotection