Generation of Proteins with Free N-Terminal Cysteine by Aminopeptidases

Journal of the American Chemical Society, Год журнала: 2022, Номер 144(47), С. 21763 - 21771

Опубликована: Ноя. 15, 2022

Efficient, site-specific, and bio-orthogonal conjugation of chemical functionalities to proteins is great utility in fundamental research as well industrial processes (e.g., the production antibody-drug conjugates immobilization enzymes for biocatalysis). A popular approach involves reacting a free N-terminal cysteine with variety electrophilic reagents. However, current methods generating cysteines have significant limitations. Herein we report novel, efficient, convenient method producing recombinant by genetically fusing Met-Pro-Cys sequence N-terminus protein interest subjecting sequential action methionine proline aminopeptidases. The resulting was site-specifically labeled at fluorescein cyclic cell-penetrating peptide through native ligation 2-cyanobenzothiazole moiety, respectively. In addition, optimal recognition Aeromonas sobria aminopeptidase determined screening combinatorial library incorporated into most efficient processing.

Язык: Английский

---

Toward a Traceless Tag for the Thiol‐Mediated Uptake of Proteins

ChemistryEurope, Год журнала: 2023, Номер 1(3)

Опубликована: Окт. 11, 2023

Abstract The emergence of thiol‐mediated uptake (TMU) as a powerful strategy to penetrate cells calls for the development practical small‐molecule TMU tools traceless delivery. Toward this goal, esters are explored here bioreversible linkers between dynamic covalent cascade exchangers accounting and protein interest (POI). method relies on α ‐aryl‐ ‐diazoamides that react with carboxylic acids POI form can be enzymatically hydrolyzed inside release native POI. A two‐step protocol is established conjugation tags Despite small number attached POIs prevent isoelectric precipitation, shown efficiently enter not only in 2D culture but also 3D spheroids mimicking deep tissue, confirming key advantage TMU. Uptake inhibition by various thiol‐reactive agents confirms participation cell‐surface thiols cell penetration, i. e., occurrence

Язык: Английский

---

Isoindolium‐Based Allenes: Reactivity Studies and Applications in Fluorescence Temperature Sensing and Cysteine Bioconjugation

Angewandte Chemie International Edition, Год журнала: 2023, Номер 62(12)

Опубликована: Янв. 21, 2023

Abstract The reaction of a series electron‐deficient isoindolium‐based allenes with sulfhydryl compounds has been studied, leading to the formation vinyl sulfides. sulfides generated could be readily converted into corresponding indanones and amines upon heating at 30–70 °C good yields up 61 %. thermal cleavage was further studied for developing temperature‐sensitive systems. Notably, novel FRET‐based fluorescent temperature sensor designed synthesized sensing 50 °C, giving 6.5‐fold blue fluorescence enhancement. Moreover, chemoselective bioconjugation cysteine‐containing peptides construction multifunctional peptide bioconjugates investigated. Thermal isoindoliums on modified 35–70 gave indanone >99 % conversion. These results indicated biocompatibility this reaction.

Язык: Английский

---

Site-specific protein labeling: Recent progress

Chinese Chemical Letters, Год журнала: 2024, Номер unknown, С. 110546 - 110546

Опубликована: Окт. 1, 2024

Язык: Английский

---

Boron enabled bioconjugation chemistries

Chemical Society Reviews, Год журнала: 2024, Номер unknown

Опубликована: Янв. 1, 2024

Organoboron compounds have given rise to a growing collection of bioconjugation reactions, with some being reversible while others yielding stable linkage. Both reaction subtypes found their unique applications in biology.

Язык: Английский

---