Photochemical cleavage of leader peptides

Bindman, Noah A.; Merkx, Remco; Koehler, Robert; Herrman, Nicholas W.C.; Donk, Wilfred A. van der

doi:10.1039/c0cc02945a

Cited by 28 publications

(19 citation statements)

References 25 publications

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“…In the last step prior to cleaving the peptides from the resin, the CuAAC ligation handles 7 or 8 (Figure 6 ) were coupled to their N-termini. The building block 7 would enable convenient removal of the leader peptide by photolysis, 38 whereas building block 8 was expected to improve the efficiency of the ligation owing to the copper ligating ability of the pyridine ligand. 39 Two ProcA2.8 precursor peptides 9 and 10 were thus synthesized by CuAAC to probe directionality of dehydration (Figure 7 A,C).…”

Section: Resultsmentioning

confidence: 99%

Mechanistic Studies on the Substrate-Tolerant Lanthipeptide Synthetase ProcM

2014

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Lanthipeptides are a class of post-translationally modified peptide natural products. They contain lanthionine (Lan) and methyllanthionine (MeLan) residues, which generate cross-links and endow the peptides with various biological activities. The mechanism of a highly substrate-tolerant lanthipeptide synthetase, ProcM, was investigated herein. We report a hybrid ligation strategy to prepare a series of substrate analogues designed to address a number of mechanistic questions regarding catalysis by ProcM. The method utilizes expressed protein ligation to generate a C-terminal thioester of the leader peptide of ProcA, the substrate of ProcM. This thioester was ligated with a cysteine derivative that resulted in an alkyne at the C-terminus of the leader peptide. This alkyne in turn was used to conjugate the leader peptides to a variety of synthetic peptides by copper-catalyzed azide–alkyne cycloaddition. Using deuterium-labeled Ser and Thr in the substrate analogues thus prepared, dehydration by ProcM was established to occur from C-to-N-terminus for two different substrates. Cyclization also occurred with a specific order, which depended on the sequence of the substrate peptides. Furthermore, using orthogonal cysteine side-chain protection in the two semisynthetic peptide substrates, we were able to rule out spontaneous non-enzymatic cyclization events to explain the very high substrate tolerance of ProcM. Finally, the enzyme was capable of exchanging protons at the α-carbon of MeLan, suggesting that ring formation could be reversible. These findings are discussed in the context of the mechanism of the substrate-tolerant ProcM, which may aid future efforts in lanthipeptide engineering.

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Section: Resultsmentioning

confidence: 99%

Mechanistic Studies on the Substrate-Tolerant Lanthipeptide Synthetase ProcM

2014

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“…Following to the N‐terminal Cys coupling, the Lys438 was deprotected under Pd‐catalysis. Subsequently, a synthesized light cleavable linker ( 2 ) (see Supporting Information) was incorporated on the free amino group (Scheme ). The reduction of the azido group and the biotin coupling were conducted in a one‐pot reaction.…”

Section: Resultsmentioning

confidence: 99%

“…and DIPEA (85 µl, 0.5 mmol, 5 eq.) in 4 ml DMF . The reaction was shaken for 20 h at rt and then washed with DMF (5x), CH 2 Cl 2 (5x), DMF (5x).…”

Section: Methodsmentioning

confidence: 99%

Semi‐synthesis of a tag‐free O‐GlcNAcylated tau protein by sequential chemoselective ligation

Schwagerus

Reimann

Despres

et al. 2016

Journal of Peptide Science

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In this paper, the first semi-synthesis of the Alzheimer-relevant tau protein carrying an O-GlcNAcylation is demonstrated by using sequential chemoselective ligation. The 52-amino acid C-terminus of tau was obtained by native chemical ligation between two synthetic peptide fragments, one carrying the O-GlcNAc moiety on Ser400, which has recently been demonstrated to inhibit tau phosphorylation and to hinder tau oligomerization, and the other equipped with a photocleavable biotin handle. After desulfurization to deliver a native alanine at the ligation junction, the N-terminal cysteine was unmasked, and the peptide was further used for expressed protein ligation to generate the full-length tau protein, which was purified by a photocleavable biotin tag. We thus provide a synthetic route to obtain a homogenous tag-free O-GlcNAcylated tau protein that can further help to elucidate the significance of posttranslational modification on the tau protein and pave the way for evaluating possible drug targets in Alzheimer's disease. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd.

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“…The design of the PCL was derived from 1‐(2‐nitrophenyl)propargyl alcohol, which was used as a photolytically cleavable linker of two peptide fragments . Nitrobenzene derivatives are appropriate linkers based on their photochemical properties and use as peptide caging groups . In order to avoid interference of deprotection with the nucleobase absorption, the wavelength for uncaging was shifted to λ =347 nm by applying the 3,4‐methoxy nitrobenzene derivative .…”

Section: Methodsmentioning

confidence: 99%

Native Chemical Ligation Directed by Photocleavable Peptide Nucleic Acid (PNA) Templates

et al. 2017

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A novel peptide-peptide ligation strategy is introduced that has the potential to provide peptide libraries of linearly or branched coupled fragments and will be suited to introduce simultaneous protein modifications at different ligation sites. Ligation is assisted by templating peptide nucleic acid (PNA) strands, and therefore, ligation specificity is solely encoded by the PNA sequence. PNA templating, in general, allows for various kinds of covalent ligation reactions. As a proof of principle, a native chemical ligation strategy was elaborated. This PNA-templated ligation includes easy on-resin procedures to couple linkers and PNA to the respective peptides, and a traceless photocleavage of the linker/PNA oligomer after the ligation step. A 4,5-dimethoxy-2-nitrobenzaldehyde-based linker that allowed the photocleavable linkage of two bio-oligomers was developed.

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Photochemical cleavage of leader peptides

Abstract: We report a photolabile linker compatible with Fmoc solid phase peptide synthesis and Cu(I)-catalyzed alkyne–azide cycloaddition that allows photochemical cleavage to afford a C-terminal peptide fragment with a native amino terminus.

Cited by 28 publications

References 25 publications

Mechanistic Studies on the Substrate-Tolerant Lanthipeptide Synthetase ProcM

Mechanistic Studies on the Substrate-Tolerant Lanthipeptide Synthetase ProcM

Semi‐synthesis of a tag‐free O‐GlcNAcylated tau protein by sequential chemoselective ligation

Native Chemical Ligation Directed by Photocleavable Peptide Nucleic Acid (PNA) Templates

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