Enantiomeric histidine-rich peptide coacervates enhance antigen delivery to T cells DOI
Ushasi Pramanik, Anirban Das, Elise M. Brown

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Sept. 14, 2024

SUMMARY Peptides and peptidomimetics that self-assemble via LLPS have recently emerged as building blocks for fabricating functional biomaterials due to their unique physicochemical properties dynamic nature. One of life’s most distinctive signatures is its selectivity chiral molecules and, date, coacervates comprised D-amino acids not been reported. Here, we demonstrate histidine-rich repeats (GHGXY) 4 (X=L/V/P) enantiomers undergo opening new avenues enhancing coacervate stability. Through a series biophysical studies, find kinetics, droplet size, fusion, encapsulation efficiency are dictated by the primary sequence. Further, these can encapsulate therapeutic cargo which then internalized endocytic mechanisms. Finally, show enhance antigen presentation CD4 + CD8 T cells resulting in robust proliferation production cytokines. Collectively, our study describes development characterization enantiomeric peptide attractive vaccine delivery vehicles with tunable properties. HIGHLIGHTS D amino acid-peptides were used first time construct phase separating Chirality does restrict or modulate other Antigen using enhances prolongs PROGRESS AND POTENTIAL self-assembly liquid-liquid separation (LLPS) result solute-rich serve biomaterials. Using repeats, this work demonstrates peptides composed entirely form coaceravtes. The kinetics bulk droplets be controlled through simple acid substitutions. coacervates, while immunologically inert, exert an adjuvanting effect leading cytokine production. materials showcased here possess high translational potential combined immunomodulators antigens against infectious diseases cancer. deliverables from will also inspire systems contribute knowledge cellular processes associated changes integral both physiology pathology.

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

62
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

28
D‐Peptide and D‐Protein Technology: Recent Advances, Challenges, and Opportunities** DOI Creative Commons
Alexander J. Lander, Yi Jin, Louis Y. P. Luk

et al.

ChemBioChem, Journal Year: 2022, Volume and Issue: 24(4)

Published: Oct. 24, 2022

Abstract Total chemical protein synthesis provides access to entire D‐protein enantiomers enabling unique applications in molecular biology, structural and bioactive compound discovery. Key enzymes involved the central dogma of biology have been prepared their D‐enantiomeric forms facilitating development mirror‐image life. Crystallization a racemic mixture L‐ high‐resolution X‐ray structures polypeptides. Additionally, D‐enantiomers drug targets can be used phage display allowing discovery non‐proteolytic D‐peptide ligands as lead candidates. This review discusses D‐proteins including synthetic challenges opportunities.

Language: Английский

Citations

44
Backbone‐Installed Split Intein‐Assisted Ligation for the Chemical Synthesis of Mirror‐Image Proteins DOI
Baochang Zhang, Yupeng Zheng, Guo‐Chao Chu

et al.

Angewandte Chemie International Edition, Journal Year: 2023, Volume and Issue: 62(33)

Published: June 26, 2023

Abstract Membrane‐associated D‐proteins are an important class of synthetic molecules needed for D‐peptide drug discovery, but their chemical synthesis using canonical ligation methods such as native is often hampered by the poor solubility constituent peptide segments. Here, we describe a B ackbone‐ I nstalled S plit ntein‐ A ssisted L igation (BISIAL) method these proteins, wherein L‐forms N‐ and C‐intein fragments unique consensus‐fast (Cfa) (i.e. L–Cfa N C ) separately installed onto two segments to be ligated via removable backbone modification. The proceeds smoothly at micromolar (μM) concentrations under strongly chaotropic conditions (8.0 M urea), subsequent removal modification groups affords desired without leaving any “ligation scar” on products. effectiveness practicality BISIAL exemplified D‐enantiomers extracellular domains T cell immunoglobulin ITIM domain (TIGIT) tropomyosin receptor kinase (TrkC). further expands protein toolkit provides practical access challenging D‐protein targets.

Language: Английский

Citations

35
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
Identification and Detection of a Peptide Biomarker and Its Enantiomer by Nanopore DOI Creative Commons
Laura Ratinho, Laurent Bacri, Bénédicte Thiébot

et al.

ACS Central Science, Journal Year: 2024, Volume and Issue: 10(6), P. 1167 - 1178

Published: May 3, 2024

Until now, no fast, low-cost, and direct technique exists to identify detect protein/peptide enantiomers, because their mass charge are identical. They essential since l- d-protein enantiomers have different biological activities due unique conformations. Enantiomers potential for diagnostic purposes several diseases or normal bodily functions but yet be utilized. This work uses an aerolysin nanopore electrical detection vasopressin l-AVP d-AVP, associated with processes pathologies. We show identification according conformations, in either native reducing conditions, using specific signature. To improve identification, we used a principal component analysis approach define the most relevant parameters identification. Finally, Monte Carlo prediction assign each event type d-AVP enantiomer.

Language: Английский

Citations

9
The Potential of Peptide-Based Inhibitors in Disrupting Protein–Protein Interactions for Targeted Cancer Therapy DOI Open Access

Ana Maria Sardinha Afonso,

Carlos Cavaleiro, Miguel A. R. B. Castanho

et al.

International Journal of Molecular Sciences, Journal Year: 2025, Volume and Issue: 26(7), P. 3117 - 3117

Published: March 28, 2025

Protein-protein interactions (PPIs) form an intricate cellular network known as the interactome, which is essential for various processes, such gene regulation, signal transduction, and metabolic pathways. The dysregulation of this has been closely linked to disease states. In cancer, these aberrant PPIs, termed oncogenic PPIs (OncoPPIs), are involved in tumour formation proliferation. Therefore, inhibition OncoPPIs becomes a strategy targeted cancer therapy. Small molecule inhibitors have dominant PPI owing their small size ability cross cell membranes. However, peptide-based emerged compelling alternatives, offering distinct advantages over inhibitors. Peptides, with larger flexible backbones, can effectively engage broad interfaces PPIs. Their high specificity, lower toxicity, ease modification make them promising candidates Over past decade, significant advancements made developing This review discusses critical aspects targeting emphasizes significance therapy, explores using therapeutic agents. It also highlights recent progress peptide design aimed at overcoming limitations therapeutics, comprehensive overview current landscape potential treatment.

Language: Английский

Citations

1
A cysteine-specific solubilizing tag strategy enables efficient chemical protein synthesis of difficult targets DOI Creative Commons
Wenchao Li,

Michael T. Jacobsen,

Claire Park

et al.

Chemical Science, Journal Year: 2024, Volume and Issue: 15(9), P. 3214 - 3222

Published: Jan. 1, 2024

We developed a new cysteine-specific solubilizing tag strategy via cysteine-conjugated succinimide. This remains stable under common native chemical ligation conditions and can be efficiently removed with palladium-based catalysts. Utilizing this approach, we synthesized two proteins containing notably difficult peptide segments: interleukin-2 (IL-2) insulin. IL-2 synthesis represents the simplest most efficient approach to date, which is enabled by synthesize ligate long segments. Additionally, T8P insulin variant, previously identified in an infant neonatal diabetes. show that exhibits reduced bioactivity (a 30-fold decrease compared standard insulin), potentially contributing onset of diabetes these patients. In summary, our work provides tool challenging opens avenues for exploring research directions understanding their biological functions.

Language: Английский

Citations

5
Engineering a Low-Immunogenic Mirror-Image VHH against Vascular Endothelial Growth Factor DOI

Keisuke Aoki,

Katsuaki Higashi,

Sakiho Oda

et al.

ACS Chemical Biology, Journal Year: 2024, Volume and Issue: 19(5), P. 1194 - 1205

Published: May 2, 2024

Immunogenicity is a major caveat of protein therapeutics. In particular, the long-term administration therapeutic agents leads to generation antidrug antibodies (ADAs), which reduce drug efficacy while eliciting adverse events. One promising solution this issue use mirror-image proteins consisting d-amino acids, are resistant proteolytic degradation in immune cells. We have recently reported chemical synthesis enantiomeric form variable domain antibody heavy chain (d-VHH). However, identifying capable binding natural ligands remains challenging. study, we developed novel screening platform identify d-VHH specific for vascular endothelial growth factor A (VEGF-A). performed two newly constructed synthetic VHH libraries displayed on T7 phage and identified sequences that effectively bound VEGF-A target (d-VEGF-A). subsequently synthesized candidate preferentially native (l-VEGF-A) with submicromolar affinity. Furthermore, immunization studies mice demonstrated elicited no ADAs, unlike its corresponding l-VHH. Our findings highlight utility development therapeutics exhibiting both reduced immunogenicity improved efficacy.

Language: Английский

Citations

5
Has AlphaFold 3 Solved the Protein Folding Problem for D-Peptides? DOI Open Access

Henry R. Childs,

Pei Zhou, Bruce R. Donald

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: March 17, 2025

Abstract Due to the favorable chemical properties of mirrored chiral centers (such as improved stability, bioavailability, and membrane permeability) computational design D-peptides targeting biological L-proteins is a valuable area research. To these structures in silico , workflow should correctly dock fold peptide while maintaining centers. The latest AlphaFold 3 (AF3) from Abramson et al. (2024) enforces strict violation penalty maintain model inputs reported have low rate only 4.4% on PoseBusters benchmark containing diverse molecules. Herein, we report results 3,255 experiments with AF3 evaluate its ability predict fold, chirality, binding pose heterochiral complexes. Despite our specifying explicit D-stereocenters, that for D-peptide binders much higher at 51% across all evaluated predictions; average accurate chance (random chirality choice, L or D, each residue). Increasing number seeds failed improve this rate. predictions exhibit incorrect folds poses, commonly oriented incorrectly L-protein pocket. Confidence metrics returned by also fail distinguish correct docking vs. high docking. We conclude poor predictor pose. Summary A crucial task protein predicting property determines structure function protein. 1 published Nature (AF3), powerful deep learning framework both bound unbound states. This architecture tuned respect centers, which are atoms (in proteins, backbone α -carbons) covalently four different species 2 . These adopt two non-superposable forms, often called “handedness,” termed (all proteins form) D (the mirror image L). exert significant influence function; changing even single residue can dramatically alter such enantioselective (e.g., antifolate resistance ) stability 4 Additionally, (small exclusively centers) many advantages compared their L-peptide counterparts, protease evasion 5 therefore therapeutically relevant modalities. vastly differing properties, an algorithm center error 0%. Although reports found explicitly specified 51%. fails Our data highlights structural prediction demonstrates Compared empirical structures, highly inaccurate when folding D-peptide:L-protein failure accurately interactions indicates more work need high-quality D-peptides.

Language: Английский

Citations

0