1,6-Thiol Addition of Trifunctional N-Alkylpyridinium for Site-selective Dual Functionalization of Proteins DOI Creative Commons
Tanja Weil, Seah Ling Kuan,

Lujuan Xu

et al.

Published: April 22, 2024

We report N-alkylpyridinium reagents for site-selective dual modification of proteins via a regioselective 1,6-thiol addition reaction. The derivatives can be syn-thesized in two reaction steps and demonstrate high labelling efficiency chemose-lectivity towards cysteine residues. This is combined with strain-promoted azide-alkyne click inverse-electron-demand Diels−Alder reactions to achieve functionalization sequential one-pot A Rho-inhibiting enzyme functionalized cancer cell-targeting peptide fluorescent dye successful vitro uptake imaging concomitant inhibition specific Rho-mediated cellular pathways. ease synthesis, fast kinetics, solubility regioselectivity make unique protein/peptide increase their functional diversities medical applications.

Language: Английский

Strain-release alkylation of Asp12 enables mutant selective targeting of K-Ras-G12D DOI Creative Commons
Qinheng Zheng, Ziyang Zhang, Keelan Z. Guiley

et al.

Nature Chemical Biology, Journal Year: 2024, Volume and Issue: 20(9), P. 1114 - 1122

Published: March 5, 2024

K-Ras is the most commonly mutated oncogene in human cancer. The recently approved non-small cell lung cancer drugs sotorasib and adagrasib covalently capture an acquired cysteine K-Ras-G12C mutation lock it a signaling-incompetent state. However, covalent inhibition of G12D, frequent particularly prevalent pancreatic ductal adenocarcinoma, has remained elusive due to lack aspartate-targeting chemistry. Here we present set malolactone-based electrophiles that exploit ring strain crosslink K-Ras-G12D at mutant aspartate form stable complexes. Structural insights from X-ray crystallography exploitation stereoelectronic requirements for attack electrophile allowed development substituted malolactone resisted by aqueous buffer but rapidly crosslinked with aspartate-12 both GDP GTP GTP-state targeting effective suppression downstream signaling, selective K-Ras-G12D-driven proliferation vitro xenograft growth mice.

Language: Английский

Citations

34
Photocatalytic Activation of Aryl(trifluoromethyl) Diazos to Carbenes for High-Resolution Protein Labeling with Red Light DOI
David C. Cabanero, Stavros K. Kariofillis, Andrew C. Johns

et al.

Journal of the American Chemical Society, Journal Year: 2024, Volume and Issue: 146(2), P. 1337 - 1345

Published: Jan. 2, 2024

State-of-the-art methods in photoproximity labeling center on the targeted generation and capture of short-lived reactive intermediates to provide a snapshot local protein environments. Diazirines are current gold standard for high-resolution proximity labeling, generating aryl(trifluoromethyl) carbenes. Here, we present method access carbenes from stable diazo source via tissue-penetrable, deep red near-infrared light (600–800 nm). The operative mechanism this activation involves Dexter energy transfer photoexcited osmium(II) photocatalysts diazo, thus revealing an carbene. preferences probe with amino acids studied, showing high reactivity toward heteroatom−H bonds. Upon synthesis biotinylated probe, studies conducted native proteins as well conjugated Os photocatalyst. Finally, demonstrate that conjugation inhibitor photocatalyst also enables selective presence spectator achieves specific membrane surface mammalian cells two-antibody photocatalytic system.

Language: Английский

Citations

22
Simultaneous Covalent Modification of K-Ras(G12D) and K-Ras(G12C) with Tunable Oxirane Electrophiles DOI
Zhongtang Yu, Xiaoqiang He, R. Wang

et al.

Journal of the American Chemical Society, Journal Year: 2023, Volume and Issue: 145(37), P. 20403 - 20411

Published: Aug. 3, 2023

Owing to their remarkable pharmaceutical properties compared those of noncovalent inhibitors, the development targeted covalent inhibitors (TCIs) has emerged as a powerful method for cancer treatment. The K-Ras mutant, which is prevalent in multiple cancers, been confirmed be crucial drug target treatment various malignancies. However, although K-Ras(G12D) mutation present up 33% mutations, no targeting have developed date. relatively weak nucleophilicity acquired aspartic acid (12D) residue may reason this. Herein, we first compound capable covalently engaging both and K-Ras(G12C) mutants. Proteome profiling revealed that this effectively conjugates with G12C G12D residues, modulating protein functions situ. These findings offer unique pathway novel dual inhibitors.

Language: Английский

Citations

29
Traceless Peptide and Protein Modification via Rational Tuning of Pyridiniums DOI
Chuan Wan, Yichi Zhang, Jinpeng Wang

et al.

Journal of the American Chemical Society, Journal Year: 2024, Volume and Issue: 146(4), P. 2624 - 2633

Published: Jan. 19, 2024

Herein, we report a versatile reaction platform for tracelessly cleavable cysteine-selective peptide/protein modification. This offers highly tunable and predictable conjugation cleavage by rationally estimating the electron effect on nucleophilic halopyridiniums. Cleavable peptide stapling, antibody conjugation, enzyme masking/de-masking, proteome labeling were achieved based this facile pyridinium-thiol-exchange protocol.

Language: Английский

Citations

11
Photoaffinity labelling with small molecules DOI

Rick A. Homan,

John D. Lapek, Christina M. Woo

et al.

Nature Reviews Methods Primers, Journal Year: 2024, Volume and Issue: 4(1)

Published: May 2, 2024

Language: Английский

Citations

10
Multiprobe Photoproximity Labeling of the EGFR Interactome in Glioblastoma Using Red-Light DOI
Feifei Tong, Weidong Zhou, Michalina Janiszewska

et al.

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

Published: March 7, 2025

Photocatalytic proximity labeling has emerged as a valuable technique for studying interactions between biomolecules in cellular context, providing precise spatiotemporal control over protein labeling. One significant advantage of these methods is their modularity, allowing the use single photocatalyst with different reactive probes to expand interactome coverage and capture diverse interactions. Despite advances, fewer have been developed using red-light excitation, limiting photoproximity more complex media such tissues animal models. Herein, we develop platform under utilizing catalyst two distinct probe types. We first design carbene based system that utilizes sulfonium diazo probes. This successfully applied on A549 cells epidermal growth factor receptor (EGFR) Cetuximab-Chlorin e6 conjugate. Benchmarking against established techniques indicates this approach performs comparably leading carbene-based methods. Next, leverage strong singlet oxygen generation (SOG) ability Chlorin establish an alternative aniline hydrazide EGFR directed chemoproteomics experiments reveal overlap system, capturing subset identified by SOG system. Finally, deploy our characterization resected human glioblastoma (GBM) tissue samples removed from locations same tumor, representing tumor's infiltrating edge its viable center, identifying several GBM specific interacting proteins may serve launch point future therapeutic campaigns.

Language: Английский

Citations

1
Evaluation of the Cytosolic Uptake of HaloTag Using a pH-Sensitive Dye DOI
JoLynn B. Giancola, Jonathan B. Grimm, Joomyung V. Jun

et al.

ACS Chemical Biology, Journal Year: 2024, Volume and Issue: 19(4), P. 908 - 915

Published: March 25, 2024

The efficient cytosolic delivery of proteins is critical for advancing novel therapeutic strategies. Current methods are severely limited by endosomal entrapment, and detection lack sophistication in tracking the fate delivered protein cargo. HaloTag, a commonly used chemical biology challenging target, an exceptional model system understanding exploiting cellular delivery. Here, we employed combinatorial strategy to direct HaloTag cytosol. We established use Virginia Orange, pH-sensitive fluorophore, Janelia Fluor 585, similar but pH-agnostic fluorogenic assay ascertain localization within human cells. Using this assay, investigated upon modification with cell-penetrating peptides, carboxyl group esterification, cotreatment endosomolytic agent. found efficacious entry two distinct methods. This study expands toolkit detecting access highlights that multiple intracellular strategies can be synergistically effect access. Moreover, poised serve as platform varied cargo into

Language: Английский

Citations

8
E3MPH16: An efficient endosomolytic peptide for intracellular protein delivery DOI
Yoshimasa Kawaguchi, Yuki Kawamura, H. Hirose

et al.

Journal of Controlled Release, Journal Year: 2024, Volume and Issue: 367, P. 877 - 891

Published: Feb. 20, 2024

Language: Английский

Citations

6
Bioreversible Anionic Cloaking Enables Intracellular Protein Delivery with Ionizable Lipid Nanoparticles DOI Creative Commons
Azmain Alamgir, Souvik Ghosal, Matthew P. DeLisa

et al.

ACS Central Science, Journal Year: 2024, Volume and Issue: 10(6), P. 1179 - 1190

Published: May 14, 2024

Protein-based therapeutics comprise a rapidly growing subset of pharmaceuticals, but enabling their delivery into cells for intracellular applications has been longstanding challenge. To overcome the barrier, we explored reversible, bioconjugation-based approach to modify surface charge protein cargos with an anionic "cloak" facilitate electrostatic complexation and lipid nanoparticle (LNP) formulations. We demonstrate that conjugation lysine-reactive sulfonated compounds can allow various using FDA-approved LNP formulations ionizable cationic DLin-MC3-DMA (MC3). apply this strategy functionally deliver RNase A cancer cell killing as well full-length antibody inhibit oncogenic β-catenin signaling. Further, show LNPs encapsulating cloaked fluorescent proteins distribute major organs in mice following systemic administration. Overall, our results point toward generalizable platform be employed wide range cargos.

Language: Английский

Citations

6
Catalytic Stereoselective 1,2-cis-Furanosylations Enabled by Enynal-Derived Copper Carbenes DOI
Bidhan Ghosh,

Adam Alber,

Chance Lander

et al.

ACS Catalysis, Journal Year: 2024, Volume and Issue: 14(2), P. 1037 - 1049

Published: Jan. 8, 2024

1,2-cis-Furanosides are present in various biomedically relevant glycosides, and their stereoselective synthesis remains a significant challenge. In this vein, we have developed approach to 1,2-cis-furanosylations using earth-abundant copper catalysis. This protocol proceeds under mild conditions at room temperature employs readily accessible benchtop stable enynal-derived furanose donors. chemistry accommodates variety of alcohols, including primary, secondary, tertiary, as well mannosyl alcohol acceptors, which been incompatible with most known methods furanosylation. The resulting 1,2-cis-furanoside products exhibit high yields anomeric selectivity both the ribose arabinose series. Furthermore, is independent C2 oxygen-protecting group configuration starting donor. Experimental evidence computational studies support our hypothesis that chelation between oxygen donor an incoming nucleophile responsible for observed 1,2-cis-stereoselectivity.

Language: Английский

Citations

5