Material-specific binding peptides empower sustainable innovations in plant health, biocatalysis, medicine and microplastic quantification

Chemical Society Reviews, Journal Year: 2024, Volume and Issue: 53(12), P. 6445 - 6510

Published: Jan. 1, 2024

Overview of natural and engineered material-binding peptides the molecular forces crucial for their (material-specific) binding to material surfaces.

Language: Английский

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Sortase-mediated labeling: Expanding frontiers in site-specific protein functionalization opens new research avenues

Current Opinion in Chemical Biology, Journal Year: 2024, Volume and Issue: 80, P. 102443 - 102443

Published: March 19, 2024

Language: Английский

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Engineered Antibodies as Cancer Radiotheranostics

Advanced Science, Journal Year: 2024, Volume and Issue: 11(30)

Published: June 14, 2024

Radiotheranostics is a rapidly growing approach in personalized medicine, merging diagnostic imaging and targeted radiotherapy to allow for the precise detection treatment of diseases, notably cancer. Radiolabeled antibodies have become indispensable tools field cancer theranostics due their high specificity affinity cancer-associated antigens, which allows accurate targeting with minimal impact on surrounding healthy tissues, enhancing therapeutic efficacy while reducing side effects, immune-modulating ability, versatility flexibility engineering conjugation. However, there are inherent limitations using as platform radiopharmaceuticals natural activities within immune system, large size preventing effective tumor penetration, relatively long half-life concerns prolonged radioactivity exposure. Antibody can solve these challenges preserving many advantages immunoglobulin framework. In this review, goal give general overview antibody design radiotheranostics. Particularly, four ways that applied enhance radioimmunoconjugates: pharmacokinetics optimization, site-specific bioconjugation, modulation Fc interactions, bispecific construct creation discussed. The radionuclide choices designed conjugates conjugation techniques future directions conjugate innovation advancement also

Language: Английский

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Substrate recognition by a peptide-aminoacyl-tRNA ligase

Proceedings of the National Academy of Sciences, Journal Year: 2025, Volume and Issue: 122(12)

Published: March 19, 2025

The continuing discovery of new peptide-aminoacyl-tRNA ligases (PEARLs) has unveiled a diverse array enzymes with the unique potential to append amino acids C terminus substrate peptides in an aminoacyl-tRNA-dependent manner. To date, PEARLs have been reported that can conjugate Cys, Ala, Trp, Gly, Leu, Asn, and Thr residues, but basis peptide aminoacyl-tRNA recognition is not known. Cell-free expression (CFE) emerged as powerful tool rapidly assay activity variants, we used technique this study investigate specificity PEARL [Formula: see text]. This enzyme adds Trp was discovered previously during genome mining for ribosomally synthesized posttranslational modified (RiPPs). remarkably tolerant changes C-terminal acid substrate, truncation replacement experiments suggest minimal sequence requirement. An AlphaFold3 model provided insights into binding interactions BhaA-Ala text] also generated predictions tRNA, ATP, Mg2+ modes were tested by site-directed mutagenesis. data several highly conserved residues recognize 3'-CCA present all tRNAs. required incorporation employed protein tag labeling eGFP, lysozyme, MBP 5-Br-Trp.

Language: Английский

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Genetic Code Expansion Approaches to Decipher the Ubiquitin Code

Chemical Reviews, Journal Year: 2024, Volume and Issue: 124(20), P. 11544 - 11584

Published: Sept. 23, 2024

The covalent attachment of Ub (ubiquitin) to target proteins (ubiquitylation) represents one the most versatile PTMs (post-translational modifications) in eukaryotic cells. Substrate modifications range from a single moiety being attached protein complex chains that can also contain Ubls (Ub-like proteins). Ubiquitylation plays pivotal roles aspects biology, and cells dedicate an orchestrated arsenal enzymes install, translate, reverse these modifications. entirety this system is coined code. Deciphering code challenging due difficulty reconstituting enzymatic machineries generating defined Ub/Ubl-protein conjugates. This Review provides comprehensive overview recent advances using GCE (genetic expansion) techniques study We highlight strategies site-specifically ubiquitylate discuss their advantages disadvantages, as well various applications. Additionally, we review potential small chemical targeting Ub/Ubls present GCE-based approaches additional layer complexity. Furthermore, explore methods rely on develop tools probe interactors offer insights into how future could help unravel complexity

Language: Английский

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A High-Throughput Cell-Free Enzyme Screening System Using Redox-Responsive Hydrogel Beads as Artificial Compartments

ACS Synthetic Biology, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 13, 2025

We have developed a rapid, simple, and high-throughput screening system for recombinant enzymes using disulfide-bonded hydrogel beads (HBs) produced via microfluidic method. These redox-responsive HBs were compatible with the biosynthesis of enzyme mutants cell-free protein synthesis, fluorescent staining through an enzymatic reaction, genetic information recovery after fluorescence-activated droplet sorting (FADS). The expression microbial transglutaminase zymogen (MTGz) synthesis cross-linking-reactivity-based product validated. Next-generation sequencing (NGS) analysis genes recovered from highly identified novel mutation sites (N25 N27) in propeptide domain. introduction these mutations allowed design engineered active MTGz, demonstrating potential as artificial compartments FADS-based selection that catalyze peptide cross-linking reactions.

Language: Английский

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Helicity‐Dependent Enzymatic Peptide Cyclization

Journal of Peptide Science, Journal Year: 2025, Volume and Issue: 31(6)

Published: April 27, 2025

ABSTRACT The secondary structure plays a crucial role in the biological activity of peptides. Various strategies have been developed to stabilize particular peptide conformations, including sequence modifications and macrocyclization approaches. Often, interplay between conformational constraint flexibility is central bioactivity. Here, we investigate how α‐helicity influences enzymatic head‐to‐tail cyclization using an engineered Sortase. We show that peptides with low helicity readily undergo intramolecular cyclization, while more rigid, helical exhibit complex behaviors cyclic dimer formation. These findings reveal increased rigidity can redirect reactions from intermolecular processes, demonstrates changes molecular guide chemical reactivity. insights advance design peptide‐derived materials, hydrogels, stimuli‐responsive probes.

Language: Английский

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Development of a Long-Acting Myeloid-Derived Growth Factor via Site-Specific PEGylation

Bioconjugate Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: April 27, 2025

Extracellular myeloid-derived growth factor (MYDGF) can improve organ repair. However, short in vivo half-life hampers its therapeutic application. Herein, we developed a long-acting MYDGF via site-specific PEGylation at C-terminus. Bacterially overexpressed human carrying C-terminal Asn-Ala-Leu tripeptide motif was first ligated with synthetic azido-functionalized Gly-Ile-Gly-Lys(N3) tetrapeptide linker catalysis of [G238 V]BmAEP1, an engineered bamboo-derived asparaginyl endopeptidase (AEP)-type peptide ligase. Thereafter, the efficiently conjugated commercially available dibenzocyclooctyne (DBCO)-functionalized linear PEG30000 copper-free click chemistry. The site-specifically PEGylated (PEG-MYDGF) retained high vitro activity and showed much longer mice compared unmodified MYDGF. In diabetic mice, PEG-MYDGF significantly promoted wound healing after subcutaneous injection. Thus, represents biologic potential. present enzymatic ligation chemistry-based approach could be applied to other proteins for conjugation various functional moieties.

Language: Английский

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Immobilizing the Endogenous 5-Hydroxytryptamine Transporter by Ligand-Directed and Proximity-Catalyzed Acyl Imidazole Chemistry for Affinity Chromatographic Analysis

Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: May 3, 2025

The sensitivity, selectivity, and accuracy of immobilized protein-based methods are critically dependent on the strategies employed for protein immobilization. Compared with random immobilization approaches, site-specific covalent have emerged as promising alternatives, offering enhanced analytical performance. However, these typically require genetic modification target to incorporate a specific tag or prior purification protein, posing significant challenges immobilizing endogenous proteins. Herein, we address limitations by using 5-hydroxytryptamine transporter (5-HTT) model system. We designed probe conjugating fluvoxamine─a ligand 5-HTT─with either fluorescent reporter aminopropyl-modified silica gel through an alkyl linker containing acyl imidazole moiety reactive group. By introducing ligand-directed (LDAI) chemistry, achieved selective labeling 5-HTT in live cells without advance. Labeling experiments confirmed high specificity favorable kinetics LDAI ligands toward 5-HTT. Chromatographic analysis revealed improvements selectivity (5.7-50.9-fold), resolution (1.2-3.0-fold), sensitivity (1.0-12.0-fold) analyzing canonical when 5-HTT, compared His-tagged Halo-tagged recombinant This approach, substituting fluvoxamine other probe, can be generalized wide range proteins, versatile platform advanced techniques.

Language: Английский

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Protein Ligases: Nature’s Gift for Protein/Peptide Synthesis

BioChem, Journal Year: 2025, Volume and Issue: 5(2), P. 11 - 11

Published: May 7, 2025

Proteins are structurally and functionally diverse biomacromolecules that serve a variety of essential activities to ensure complex biological homeostasis. The desire elucidate enhance these functions has been at the forefront research for many decades. However, generating active proteins via recombinant expression or through chemical total synthesis each limitations in terms yield functionality. Nature provided solution this problem evolving protein ligases catalyze formation amide bonds between peptides/proteins, which can be exploited by engineers develop robust functional proteins. Here, we summarize biochemical mechanisms applications multiple cysteine-based ligases, especially focusing on how they have utilized therapeutics engineering, as well inspired chemists efficient methodologies (e.g., native ligation).

Language: Английский

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