Biscysteine-Bearing Peptide Probes To Reveal Extracellular Thiol–Disulfide Exchange Reactions Promoting Cellular Uptake

In recent years, delivery systems based on the incorporation of thiols/disulfides have been extensively explored to promote the intracellular delivery of biological cargoes. However, it remains unclear about the detailed processes of thiol–disulfide exchanges taking place on the cell surface and how...

Full description

Saved in:
Bibliographic Details
Published in:Analytical chemistry (Washington) 2017-08, Vol.89 (16), p.8501-8508
Main Authors: Li, Tao, Gao, Wei, Liang, Jingjing, Zha, Mirao, Chen, Yaqi, Zhao, Yibing, Wu, Chuanliu
Format: Article
Language:English
Subjects:
Cell surface
Cells
Chemical bonds
Chemical reactions
Chemistry
Cysteine
Disulfide bonds
Disulfides
Exchanging
Glutathione
Glycine
Information processing
Intracellular
Peptides
Probes
Protein thiols
Proteins
Thiols
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:In recent years, delivery systems based on the incorporation of thiols/disulfides have been extensively explored to promote the intracellular delivery of biological cargoes. However, it remains unclear about the detailed processes of thiol–disulfide exchanges taking place on the cell surface and how the exchange reactions promote the cellular uptake of cargoes bearing thiols or disulfide bonds. In this work, we report the rational design of biscysteine motif-containing peptide probes with substantially different ring-closing property and how these peptide probes were employed to explore the thiol–disulfide exchanges on the cell surface. Our results show that extensive thiol–disulfide exchanges between peptides and exofacial protein thiols/disulfides are involved in the cellular uptake of these peptide probes, and importantly glutathione (GSH) exported from the cytosols participates extensively in the exchange reactions. Cysteine−glycine−cysteine (CGC)-containing peptide probes can be more efficiently taken up by cells compared to other probes, and we suggested that the driving force for the superior cellular uptake arises from very likely the unique propensity of the CGC motif in forming doubly bridged disulfide bonds with exofacial proteins. Our probe-based strategy provides firsthand information on the detailed processes of the exchange reactions, which would be of great benefit to the development of delivery systems based on the extracellular thiol–disulfide exchanges for intracellular delivery of biologics.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.7b02084