Selective Activation of Peptide‐Thioester Precursors for Templated Native Chemical Ligations DOI Creative Commons
Paul Spaltenstein,

Riley J. Giesler,

S. Scherer

et al.

Angewandte Chemie International Edition, Journal Year: 2024, Volume and Issue: 64(1)

Published: Aug. 29, 2024

Chemical protein synthesis enables access to proteins that would otherwise be difficult or impossible obtain with traditional means such as recombinant expression. Chemoselective ligations provide the ability join peptide segments prepared by solid-phase synthesis. While native chemical ligation (NCL) is widely used, it limited need for C-terminal thioesters suitable reaction kinetics, properly placed Cys thiolated derivatives, and segment solubility at low mM concentrations. Moreover, repetitive purifications isolate ligated products are often yield-sapping, hampering efficiency progress. In this work, we demonstrate use of Controlled Activation Peptides Templated NCL (CAPTN). This traceless multi-segment templated approach permits one-pot harnessing selective thioester activation orthogonal conjugation chemistries favor formation full-length product while minimizing side reactions. Importantly, CAPTN provides kinetic enhancements allowing sterically hindered junctions Additionally, removes intermediate purification. We report two E. coli ribosomal subunits S16 S17 enabled tools described herein. anticipate will expedite valuable expand on approaches

Language: Английский

Recent advances in chemical protein synthesis: method developments and biological applications DOI
Suwei Dong, Ji‐Shen Zheng, Yiming Li

et al.

Science China Chemistry, Journal Year: 2024, Volume and Issue: 67(4), P. 1060 - 1096

Published: March 12, 2024

Language: Английский

Citations

65
Three Rounds of Stability-Guided Optimization and Systematical Evaluation of Oncolytic Peptide LTX-315 DOI

Xing‐Yan Fu,

Hao Yin,

Xi‐Tong Chen

et al.

Journal of Medicinal Chemistry, Journal Year: 2024, Volume and Issue: 67(5), P. 3885 - 3908

Published: Jan. 26, 2024

Oncolytic peptides represent promising novel candidates for anticancer treatments. In our efforts to develop oncolytic possessing both high protease stability and durable efficiency, three rounds of optimization were conducted on the first-in-class peptide LTX-315. The robust synthetic method, in vitro vivo activity, mechanism investigated. D-type represented by FXY-12 possessed significantly improved proteolytic sustained efficiency. Strikingly, hybrid FXY-30, containing one two camptothecin moieties, exhibited most potent activities. explorations indicated that FXY-30 rapid membranolytic effects induced severe DNA double-strand breaks trigger cell apoptosis. Collectively, this study not only established strategies improve potential but also provided valuable references future development peptides-based chemotherapeutics.

Language: Английский

Citations

30
Mirror-image protein and peptide drug discovery through mirror-image phage display DOI
Yun‐Kun Qi, Ji‐Shen Zheng, Lei Liu

et al.

Chem, Journal Year: 2024, Volume and Issue: 10(8), P. 2390 - 2407

Published: July 5, 2024

Language: Английский

Citations

29
Mechanically Triggered Protein Desulfurization DOI
Dongyang Han, Yan Cui, Xiangyu Deng

et al.

Journal of the American Chemical Society, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 23, 2025

The technology of native chemical ligation and postligation desulfurization has greatly expanded the scope modern protein synthesis. Here, we report that ultrasonic energy can trigger robust clean desulfurization, developed an ultrasound-induced (USID) strategy is simple to use generally applicable peptides proteins. USID involves a cleaning bath easy-to-use easy-to-remove sonosensitizer, titanium dioxide. It features mild convenient reaction conditions excellent functional group compatibility, e.g., with thiazolidine (Thz) serotonin, which are sensitive other strategies. robust: without reoptimizing conditions, same procedure be used for broad range proteins one or more sulfhydryl groups, even in multi-hundred-milligram scale reactions. utility was demonstrated by one-pot synthesis bioactive cyclopeptides such as Cycloleonuripeptide E Segetalin F, well convergent functionally important histone H3.5 using Thz temporary protecting group. A mechanistic investigation indicated proceeds via radical-based mechanism promoted low-frequency low-intensity ultrasonication. Overall, our work introduces mechanically triggered approach potential become method general both academic industrial laboratories.

Language: Английский

Citations

2
L‐Glycosidase‐Cleavable Natural Glycans Facilitate the Chemical Synthesis of Correctly Folded Disulfide‐Bonded D‐Proteins DOI
Weiwei Shi, Tongyue Wang, Ziyi Yang

et al.

Angewandte Chemie International Edition, Journal Year: 2024, Volume and Issue: 63(9)

Published: Jan. 9, 2024

D-peptide ligands can be screened for therapeutic potency and enzymatic stability using synthetic mirror-image proteins (D-proteins), but efficient acquisition of these D-proteins hampered by the need to accomplish their in vitro folding, which often requires formation correctly linked disulfide bonds. Here, we report finding that temporary installation natural O-linked-β-N-acetyl-D-glucosamine (O-GlcNAc) groups onto selected D-serine or D-threonine residues disulfide-bonded facilitate folding vitro, glycosyl completely removed from folded afford desired chirally inverted D-protein targets naturally occurring O-GlcNAcase. This approach enabled chemical syntheses several important difficult-to-fold incorporating bonds including tumor necrosis factor alpha (D-TNFα) homotrimer receptor-binding domain Omicron spike protein (D-RBD). Our work establishes use O-GlcNAc synthesis proves bearing good substrates

Language: Английский

Citations

13
Chemical Synthesis of Human Proteoforms and Application in Biomedicine DOI Creative Commons
Huasong Ai, Man Pan, Lei Liu

et al.

ACS Central Science, Journal Year: 2024, Volume and Issue: 10(8), P. 1442 - 1459

Published: July 22, 2024

Limited understanding of human proteoforms with complex posttranslational modifications and the underlying mechanisms poses a major obstacle to research on health disease. This Outlook discusses opportunities challenges

Language: Английский

Citations

12
Enhanced native chemical ligation by peptide conjugation in trifluoroacetic acid DOI Creative Commons
Dong‐Liang Huang,

Wu-Chen Guo,

Wei-Wei Shi

et al.

Science Advances, Journal Year: 2024, Volume and Issue: 10(29)

Published: July 17, 2024

Chemical ligation of peptides is increasingly used to generate proteins not readily accessible by recombinant approaches. However, a robust method ligate "difficult" remains be developed. Here, we report an enhanced native chemical strategy mediated peptide conjugation in trifluoroacetic acid (TFA). The between carboxyl-terminal thiosalicylaldehyde thioester and 1,3-dithiol-containing TFA proceeds rapidly form thioacetal-linked intermediate, which converted into the desired amide bond product through simple postligation treatment. effectiveness practicality was demonstrated successful synthesis several challenging proteins, including SARS-CoV-2 transmembrane Envelope (E) protein nanobodies. Because ability dissolve virtually all prevent formation unreactive structures, expected open new opportunities for synthesizing families particularly those with aggregable or colloidal segments.

Language: Английский

Citations

10
Efficient Delivery of Oncolytic Peptide LTX-315 by ZIF-8: pH-Responsive Release, Improved Stability, and Reduced Hemolysis DOI

Xin‐Qi Chen,

Sheng Cui, Yuzhen Chen

et al.

Molecular Pharmaceutics, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 6, 2025

The first-in-class oncolytic peptide LTX-315 has exhibited positive anticancer responses in multiple phase I/II clinical trials. Nevertheless, the linear suffers from poor proteolytic stability and undesired toxicity, especially hemolysis, which may limit its widespread applications. Except for direct structural modifications, drug delivery systems (DDSs) are expected to protect degradation shield hemolytic properties. Therefore, zeolitic imidazolate framework (ZIF-8)-based nanoparticles (NPs) were constructed with a high encapsulation rate of 59.9%, utilizing biomineralized "one-pot method" an aqueous system. release LTX-315, vitro potency, serum stability, durability, antimigration activity, hemolysis effect, subcellular localization, membrane disruption/permeation effects LTX-315@ZIF-8 NPs investigated. potent cytotoxicity against cancer cells. experiment time-inhibition curve assay indicated that ZIF-8 could effectively improve prolong duration action, enhance cytostatic potency. Especially, not only attenuated toxicity but also achieved pH-responsive LTX-315. mechanism investigation possessed membranolytic activity reduced mitochondrial potential trigger cell death. Collectively, this paper established robust strategy reduce properties provided reliable reference future peptides.

Language: Английский

Citations

1
An Enzyme‐Cleavable Solubilizing‐Tag Facilitates the Chemical Synthesis of Mirror‐Image Proteins DOI
Yupeng Zheng, Baochang Zhang, Weiwei Shi

et al.

Angewandte Chemie International Edition, Journal Year: 2024, Volume and Issue: 63(14)

Published: Feb. 8, 2024

Abstract Mirror‐image proteins (D‐proteins) are useful in biomedical research for purposes such as mirror‐image screening D‐peptide drug discovery, but the chemical synthesis of many D‐proteins is often low yielding due to poor solubility or aggregation their constituent peptide segments. Here, we report a Lys‐C protease‐cleavable solubilizing tag and its use synthesize difficult‐to‐obtain D‐proteins. Our easily installed onto multiple amino acids D Lys, Ser, Thr, and/or N‐terminal acid hydrophobic D‐peptides, impervious various reaction conditions, synthesis, ligation, desulfurization, transition metal‐mediated deprotection, yet can be completely removed by protease under denaturing conditions give desired D‐protein. The efficacy practicality new method were exemplified two challenging D‐proteins: D‐enantiomers programmed cell death protein 1 IgV domain SARS‐CoV‐2 envelope protein, high yield. This work demonstrates that enzymatic cleavage tags feasible, thus paving way production more

Language: Английский

Citations

8
Design and synthesis of TH19P01-Camptothecin based hybrid peptides inducing effective anticancer responses on sortilin positive cancer cells DOI
Yajie Li,

Chang-Bo Fang,

Shu‐shu Wang

et al.

Bioorganic & Medicinal Chemistry, Journal Year: 2024, Volume and Issue: 111, P. 117869 - 117869

Published: Aug. 3, 2024

Language: Английский

Citations

6