NanoClick: A High Throughput, Target-Agnostic Peptide Cell Permeability Assay.
Quick Facts
What This Study Found
NanoClick measures the cumulative cytosolic exposure of a peptide in a concentration-dependent manner. It combines two technologies: in-cell click chemistry (where a chemical tag on the peptide reacts with a partner inside the cell) and NanoBRET (a light signal that only fires when the peptide reaches the cytoplasm).
The assay was validated using known cell-penetrating peptides and correlated with actual cellular activity using a p53/MDM2 model system (a well-studied protein-protein interaction in cancer). This confirms NanoClick measures functionally relevant cell penetration.
With minimal changes to the peptide sequence, NanoClick can detect entry via different mechanisms: endocytosis, direct membrane translocation, or passive permeability. This versatility is important because different peptides enter cells by different routes.
Key Numbers
Validated with known CPPs; correlated with p53/MDM2 system; measures endocytosis, translocation, passive permeability
How They Did This
Researchers developed the NanoClick assay using HeLa cells expressing NanoBRET components. They validated it with known cell-penetrating peptides and a p53/MDM2 inhibitor peptide system. The assay uses click chemistry-compatible tags added to test peptides with minimal sequence disruption. High-throughput screening capability was demonstrated.
Why This Research Matters
Macrocyclic peptides can target protein-protein interactions that small molecules cannot reach, but they must get inside cells to work. The lack of a high-throughput permeability assay has been a critical bottleneck. NanoClick fills this gap and could accelerate peptide drug discovery significantly.
What This Study Doesn't Tell Us
The assay requires adding click chemistry tags to peptides, which could subtly alter their properties. It was demonstrated primarily with cyclic peptides; performance with other peptide types may vary. The assay measures cytosolic exposure but not subcellular localization. It was validated in one cell line (HeLa).
Trust & Context
- Original Title:
- NanoClick: A High Throughput, Target-Agnostic Peptide Cell Permeability Assay.
- Published In:
- ACS chemical biology, 16(2), 293-309 (2021)
- Authors:
- Peier, Andrea, Ge, Lan, Boyer, Nicolas, Frost, John, Duggal, Ruchia, Biswas, Kaustav, Edmondson, Scott, Hermes, Jeffrey D, Yan, Lin, Zimprich, Chad, Sadruddin, Ahmad, Kristal Kaan, Hung Yi, Chandramohan, Arun, Brown, Christopher J, Thean, Dawn, Lee, Xue Er, Yuen, Tsz Ying, Ferrer-Gago, Fernando J, Johannes, Charles W, Lane, David P, Sherborne, Brad, Corona, Cesear, Robers, Matthew B, Sawyer, Tomi K, Partridge, Anthony W
- Database ID:
- RPEP-05676
Evidence Hierarchy
Read More on RethinkPeptides
Cite This Study
https://rethinkpeptides.com/research/RPEP-05676APA
Peier, Andrea; Ge, Lan; Boyer, Nicolas; Frost, John; Duggal, Ruchia; Biswas, Kaustav; Edmondson, Scott; Hermes, Jeffrey D; Yan, Lin; Zimprich, Chad; Sadruddin, Ahmad; Kristal Kaan, Hung Yi; Chandramohan, Arun; Brown, Christopher J; Thean, Dawn; Lee, Xue Er; Yuen, Tsz Ying; Ferrer-Gago, Fernando J; Johannes, Charles W; Lane, David P; Sherborne, Brad; Corona, Cesear; Robers, Matthew B; Sawyer, Tomi K; Partridge, Anthony W. (2021). NanoClick: A High Throughput, Target-Agnostic Peptide Cell Permeability Assay.. ACS chemical biology, 16(2), 293-309. https://doi.org/10.1021/acschembio.0c00804
MLA
Peier, Andrea, et al. "NanoClick: A High Throughput, Target-Agnostic Peptide Cell Permeability Assay.." ACS chemical biology, 2021. https://doi.org/10.1021/acschembio.0c00804
RethinkPeptides
RethinkPeptides Research Database. "NanoClick: A High Throughput, Target-Agnostic Peptide Cell P..." RPEP-05676. Retrieved from https://rethinkpeptides.com/research/peier-2021-nanoclick-a-high-throughput
Access the Original Study
Study data sourced from PubMed, a service of the U.S. National Library of Medicine, National Institutes of Health.
This study breakdown was produced by the RethinkPeptides research team. We analyze and report published research findings without making health recommendations. All interpretations are based solely on the published abstract and study data.