Chemical Synthesis of Activity‐Based E2‐Ubiquitin Probes for the Structural Analysis of E3 Ligase‐Catalyzed Transthiolation

Liang, Lujun; Chu, Guo‐Chao; Qu, Qian; Zuo, Chong; Mao, Junxiong; Zheng, Qin; Chen, Jingnan; Meng, Xianbin; Jing, Yangwode; Deng, Haiteng; Li, Yiming; Liu, Lei

doi:10.1002/anie.202105870

Cited by 56 publications

(27 citation statements)

References 70 publications

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“…17 This method avoids the need for strong oxidizing conditions, enabling broader compatibility with common synthetic peptide functionalities, and has been widely adopted in a broad range of applications. [20][21][22][23][24][25][26][27][28][29][30][31][32][33][34][35][36][37] The increased use of C-terminal α-hydrazides in peptide chemistry creates a demand for robust synthetic tools for accessing these moieties. A number of methods have been published to allow access to C-terminal hydrazides on both synthetic 15,16,38 and expressed 15,39,40 peptides and proteins.…”

Section: Introductionmentioning

confidence: 99%

A Shelf Stable Fmoc Hydrazine Resin for the Synthesis of Peptide Hydrazides

Bird

Dawson

2022

Preprint

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C-terminal hydrazides are an important class of synthetic peptides with an ever expanding scope of applications, but their widespread application for chemical protein synthesis has been hampered due to the lack of stable resin linkers for synthesis of longer and more challenging peptide hydrazide fragments. We present a practical method for the regeneration, loading, and storage of trityl-chloride resins for the production of hydrazide containing peptides, leveraging 9-fluorenylmethyl carbazate. We show that these resins are extremely stable under several common resin storage conditions. The application of these resins to solid phase peptide synthesis (SPPS) is demonstrated through the synthesis of the 40-mer GLP-1R agonist peptide “P5”. These studies support the broad utility of Fmoc-NHNH-Trt resins for SPPS of C-terminal hydrazide peptides.

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Section: Introductionmentioning

confidence: 99%

A Shelf Stable Fmoc Hydrazine Resin for the Synthesis of Peptide Hydrazides

Bird

Dawson

2022

Preprint

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“…19 For instance, homogeneously modified histones prepared by chemical protein synthesis allow the examination of direct or indirect PTM interplays. 14,[20][21][22][23][24][25][26] Protein chemistry also enables the flexible alteration of protein at the histone and/or modifications for comprehensive mechanistic investigation. [27][28][29][30][31][32][33][34][35][36][37][38][39] Moreover, mechanism-based chemical probes can be used for trapping specific reaction states of the PTM enzymes for structural determination.…”

Section: Introductionmentioning

confidence: 99%

“…In this context, chemical methods as a complement to traditional biological technologies have been developed in the past decade to help the study of the regulation of eukaryotic chromatin by histone PTMs 19 . For instance, homogeneously modified histones prepared by chemical protein synthesis allow the examination of direct or indirect PTM interplays 14,20–26 . Protein chemistry also enables the flexible alteration of protein at the histone and/or modifications for comprehensive mechanistic investigation 27–39 .…”

Section: Introductionmentioning

confidence: 99%

Chemical methods for studying the crosstalk between histone H2B ubiquitylation and H3 methylation

Peng

2021

Journal of Peptide Science

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The reversible and dynamic post‐translational modifications (PTMs) of histones in eukaryotic chromatin are intimately connected to cell development and gene function, and abnormal regulation of PTMs can result in cancer and neurodegenerative diseases. Specific combinations of these modifications are mediated by a series of chromatin proteins that write, erase, and read the “histone codes,” but mechanistic studies of the precise biochemical and structural relationships between different sets of modifications and their effects on chromatin function constitute a unique challenge to canonical biochemical approaches. In the past decade, the development and application of chemical methods for investigating histone PTM crosstalks has received considerable attention in the field of chemical biology. In this review, taking the functional crosstalk between H2B ubiquitylation at Lys120 (H2BK120ub) and H3 methylation at Lys79 (H3K79me) as a typical example, we survey recent developments of different chemical methods, in particular, protein synthetic chemistry and protein‐based chemical probes, for studying the mechanism of the functional crosstalks of histone PTMs.

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“…12,14,16,17,18,19,20, and 21 on TRPV1 were evaluated by the patch-clamp technology. Linear peptide 1 was used as the negative control, and capsaicin (Cap), a specific agonist of TRPV1, was used as a positive control.…”

mentioning

confidence: 99%

DIC/Oxyma‐based accelerated synthesis and oxidative folding studies of centipede toxin RhTx

Chen

Wang

et al. 2021

Journal of Peptide Science

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Coupling reagents play crucial roles in the iterative construction of amide bonds for the synthesis of peptides and peptide-based derivatives. The novel DIC/Oxyma condensation system featured with the low risk of explosion displayed remarkable abilities to inhibit racemization, along with efficient coupling efficiency in both manual and automated syntheses. Nevertheless, an ideal reaction molar ratio in DIC/Oxyma condensation system and the moderate reaction temperature by manual synthesis remain to be further investigated. Herein, the synthetic efficiencies of different reaction ratios between DIC and Oxyma under moderate reaction temperature were systematically evaluated. The robustness and efficiency of DIC/Oxyma condensation system are validated by the rapid synthesis of linear centipede toxin RhTx. Different folding strategies were applied for the construction of disulfide bridges in RhTx, which was further confirmed in assays of circular dichroism and patch-clamp electrophysiology evaluation. This work establishes the DIC/Oxyma-based accelerated synthesis of peptides under moderate condensation conditions, which is especially useful for the manual synthesis of peptides. Besides, the strategy presented here provides robust technical supports for the large-scale synthesis and oxidative folding of RhTx.

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Chemical Synthesis of Activity‐Based E2‐Ubiquitin Probes for the Structural Analysis of E3 Ligase‐Catalyzed Transthiolation

Cited by 56 publications

References 70 publications

A Shelf Stable Fmoc Hydrazine Resin for the Synthesis of Peptide Hydrazides

A Shelf Stable Fmoc Hydrazine Resin for the Synthesis of Peptide Hydrazides

Chemical methods for studying the crosstalk between histone H2B ubiquitylation and H3 methylation

DIC/Oxyma‐based accelerated synthesis and oxidative folding studies of centipede toxin RhTx

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