Fast and Stable N-Terminal Cysteine Modification through Thiazolidino Boronate Mediated Acyl Transfer
- 17 August 2020
- journal article
- research article
- Published by Wiley in Angewandte Chemie-International Edition
- Vol. 59 (34), 14246-14250
- https://doi.org/10.1002/anie.202000837
Abstract
We report a novel conjugation of N-terminal cysteines (NCys) that proceeds with fast kinetics and exquisite selectivity, thereby enabling facile modification of NCys-bearing proteins in complex biological milieu. This new NCys conjugation proceeds via a thiazolidine boronate (TzB) intermediate that results from fast (k(2): approximate to 5000 m(-1) s(-1)) and reversible conjugation of NCys with 2-formylphenylboronic acid (FPBA). We designed a FPBA derivative that upon TzB formation elicits intramolecular acyl transfer to give N-acyl thiazolidines. In contrast to the quick hydrolysis of TzB, the N-acylated thiazolidines exhibit robust stability under physiologic conditions. The utility of the TzB-mediated NCys conjugation is demonstrated by rapid and non-disruptive labeling of two enzymes. Furthermore, applying this chemistry to bacteriophage allows facile chemical modification of phage libraries, which greatly expands the chemical space amenable to phage display.Funding Information
- National Institute of General Medical Sciences (GM102735)
- Directorate for Mathematical and Physical Sciences (CHE-1904874)
This publication has 28 references indexed in Scilit:
- A Biocompatible Condensation Reaction for the Labeling of Terminal Cysteine Residues on ProteinsAngewandte Chemie, 2009
- A Biocompatible Condensation Reaction for the Labeling of Terminal Cysteine Residues on ProteinsAngewandte Chemie, 2009
- Phage-encoded combinatorial chemical libraries based on bicyclic peptidesNature Chemical Biology, 2009
- HaloTag: A Novel Protein Labeling Technology for Cell Imaging and Protein AnalysisACS Chemical Biology, 2008
- An Engineered Protein Tag for Multiprotein Labeling in Living CellsCell Chemical Biology, 2008
- Sortase-Mediated Protein Ligation: A New Method for Protein EngineeringJournal of the American Chemical Society, 2004
- A general method for the covalent labeling of fusion proteins with small molecules in vivoNature Biotechnology, 2002
- Stereospecific Pseudoproline Ligation of N-Terminal Serine, Threonine, or Cysteine-Containing Unprotected PeptidesJournal of the American Chemical Society, 1999
- Synthesis of Proteins by Native Chemical LigationScience, 1994
- Neighboring-group participation of aldehydes and ketones in ester hydrolysis. Mechanism of hydrolysis of O-acetylsalicylaldehydeJournal of the American Chemical Society, 1978