One-pot multi-segment condensation strategies for chemical protein synthesis

Zuo, Chong; Zhang, Baochang; Yan, Bingjia; Zheng, Ji‐Min

doi:10.1039/c8ob02610f

Cited by 39 publications

(25 citation statements)

References 114 publications

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“…In turn, this requires a thioester to be generated at each round of ligation, which is a nontrivial feat. Although the literature is rich with sequential ligations schemes that use different peptide chemistries, the challenge of performing NCL/EPL solely with recombinant fragments has yet to be met. Herein, we have introduced a broadly applicable approach that offers a path to this goal because now ligation can be performed with a hydrazide‐containing fragment that can later be turned into a thioester for the next ligation step .…”

Section: Resultsmentioning

confidence: 99%

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Efficient Generation of Hydrazides in Proteins by RadA Split Intein

Ekanayake

Santoleri

et al. 2019

ChemBioChem

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Protein C‐terminal hydrazides are useful for bioconjugation and construction of proteins from multiple fragments through native chemical ligation. To generate C‐terminal hydrazides in proteins, an efficient intein‐based preparation method has been developed by using thiols and hydrazine to accelerate the formation of the transient thioester intermediate and subsequent hydrazinolysis. This approach not only increases the yield, but also improves biocompatibility. The scope of the method has been expanded by employing Pyrococcus horikoshii RadA split intein, which can accommodate a broad range of extein residues before the site of cleavage. The use of split RadA minimizes premature intein N cleavage in vivo and offers control over the initiation of the intein N cleavage reaction. It is expected that this versatile preparation method will expand the utilization of protein C‐terminal hydrazides in protein preparation and modification.

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Section: Resultsmentioning

confidence: 99%

“…This could be realized by using our method to generate protein fragments with an N‐terminal cysteine and a C‐terminal hydrazide. Such fragments could then be joined by NCL in any order desired –…”

Section: Resultsmentioning

confidence: 99%

Efficient Generation of Hydrazides in Proteins by RadA Split Intein

Ekanayake

Santoleri

et al. 2019

ChemBioChem

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“…Finally, to confirm the applicability of our methods to the synthesis of peptides containing sequences difficult to obtain chemically, we synthesized the amyloid β‐protein 1–42 (Aβ42), which contained an extremely hydrophobic sequence in its C‐terminal region, by combining the NCL and desulfurization strategy with the use of our solubilizing system. As determined from the data reported in Figure , [Cys21]Aβ42, containing the solubilizing tag at Glu22 ( 13 ), was synthesized by means of NCL with Aβ(1–20)‐MeNbz‐(Lys) 3 ‐NH 2 ( 11 ) and Cys21‐Glu22[Dbz‐Cys(Trt‐K 5 )‐NH 2 ]‐Aβ(23–42) ( 12 ).…”

Section: Figurementioning

confidence: 99%

Solubilizing Trityl‐Type Tag To Synthesize Asx/Glx‐Containing Peptides

2019

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A novel solubilizing tag system for Asp/Asn/Glu/Gln‐containing peptides is described. In this method, an Asp/Glu[Dbz‐Cys‐NH2]‐containing peptide (Dbz: 3,4‐diaminobenzoic acid) is first synthesized through fluorenylmethyloxycarbonyl (Fmoc) solid‐phase peptide synthesis. The solubilizing moiety containing an oligo‐Lys group is then attached to the peptide in hexafluoroisopropanol through a trityl anchor to afford a hydrophilic tagged peptide. To detach the solubilizing tag, the Dbz moiety of the tagged peptide is activated with NaNO2, and the Asp/Asn/Glu/Gln‐containing peptide is obtained through hydrolysis or ammonolysis. This synthetic approach proved to be compatible with native chemical ligation, and amyloid β‐protein 1–42 was successfully synthesized by the solubilizing‐tag‐aided native chemical ligation–desulfurization method.

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“…The fieldo fb ioorganic chemistry and chemicalb iology has recently witnessed ab oom in peptidel igation and conjugation strategies, [1] along with the rise of the bioorthogonality concept. [2] Developments include tandem,o ne-pot and solidphase schemes, [3,4] and methodologies are pushed up to the limits of synthetic viability.T he use of precisely addressable transacylation systems, [5][6][7] applied in fine-tuned combinations for the sequential, convergento ri terative assembly of polypeptidem olecules, [8][9][10] demonstrates the attained level of sophistication. The role of the native(-type) chemical ligation method(s) [11][12][13][14] confirms the particulard esire for the original amide bond, ubiquitous in life and nature,w ith specialf ocus towardsconceptual advances in amidation tactics.…”

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confidence: 99%

Untapped Opportunities of Resin‐to‐Resin Transfer Reactions (RRTR) for the Convergent Assembly of Multivalent Peptide Conjugates

Verzele

García

Madder

2020

Chemistry A European J

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Handling of the individual fragments remains a bottleneck in the convergent assembly of peptides. Overlooked since the emergence of ligation chemistries during the past two decades, so‐called resin‐to‐resin transfer reactions (RRTR) are here described as a strategic shortcut in this context. Condensation of the involved moieties at an acceptor resin is facilitated by shuttling peptide segments directly from a donor resin in a one‐pot fashion. The straightforward synthesis of a sterically constrained 13‐mer peptidosteroid model illustrates the utility of this approach, presenting the first successful application of the RRTR methodology in the field of multivalent design and bioconjugation. Relying on established procedures to generate, monitor and isolate intermediates and products, the solid‐phase nature of the entire strategy allows for the fast construction of polypeptide adducts and libraries thereof. As such, a rejuvenated use and new opportunities for RRTR are reported.

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One-pot multi-segment condensation strategies for chemical protein synthesis

Abstract: This paper describes recent advances of one-pot multi-segment condensation strategies based on kinetically controlled strategies and/or protecting group-removal strategies in chemical protein synthesis.

Cited by 39 publications

References 114 publications

Efficient Generation of Hydrazides in Proteins by RadA Split Intein

Efficient Generation of Hydrazides in Proteins by RadA Split Intein

Solubilizing Trityl‐Type Tag To Synthesize Asx/Glx‐Containing Peptides

Untapped Opportunities of Resin‐to‐Resin Transfer Reactions (RRTR) for the Convergent Assembly of Multivalent Peptide Conjugates

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