Journal
BIOORGANIC & MEDICINAL CHEMISTRY
Volume 30, Issue -, Pages -Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.bmc.2020.115947
Keywords
-
Funding
- MEXT, Japan [18 K14334, 19H05764]
- Japan Science and Technology Agency (JST) ERATO Grant [JPMJER1802]
- [17H06348]
- MRC [MC_UP_1201/15] Funding Source: UKRI
- Grants-in-Aid for Scientific Research [19H05764] Funding Source: KAKEN
Ask authors/readers for more resources
In this study, a new reactive peptide tag/probe pair system for site-specific covalent protein labeling was developed. The method relies on the recognition-driven reaction of a peptide tag and a molecular probe. The reactive profile and site-specificity of this method were characterized using model peptides and proteins in vitro, and the general utility for production of a nanobody-chemical probe conjugate without compromising its binding ability was demonstrated.
The ability to incorporate a desired functionality into proteins of interest in a site-specific manner can provide powerful tools for investigating biological systems and creating therapeutic conjugates. However, there are not any universal methods that can be applied to all proteins, and it is thus important to explore the chemical strategy for protein modification. In this paper, we developed a new reactive peptide tag/probe pair system for site-specific covalent protein labeling. This method relies on the recognition-driven reaction of a peptide tag and a molecular probe, which comprises the lysine-containing short histidine tag (KH6 or H6K) and a binuclear nickel (II)- nitrilotriacetic acid (Ni2+-NTA) complex probe containing a lysine-reactive N-acyl-N-alkyl sulfonamide (NASA) group. The selective interaction of the His-tag and Ni2+-NTA propeles a rapid nucleophilic reaction between a lysine residue of the tag and the electmphilic NASA group of the probe by the proximity effect, resulting in the tag-site-specific functionalization of proteins. We characterized the reactive profile and site-specificity of this method using model peptides and proteins in vitro, and demonstrated the general utility for production of a nanobody-chemical probe conjugate without compromising its binding ability.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available