Enzymatic N-terminal Addition of Noncanonical Amino Acids to Peptides and Proteins

Creators: Connor, Rebecca E.; Piatkov, Konstantin; Varshavsky, Alexander; Tirrell, David A.

Abstract

A chromatographic assay was used to identify ten noncanonical amino acids that can be appended to the N terminus of peptides and proteins by the L,F-transferase of Escherichia coli. A model protein substrate, E. coli dihydrofolate reductase, was modified with p-ethynylphenylalanine and conjugated to azide–biotin and azide–polyethylene glycol–fluorescein probes (see scheme).

Additional Information

© 2008 WILEY-VCH Verlag GmbH & Co. Received: October 10, 2007. Published online on January 18, 2008. This work was supported by NIH grant GM62523 (to D.A.T.), NIH grant DK39520 (to A.V.), the ARO-sponsored Institute for Collaborative Biotechnologies, and the Beckman Institute at Caltech. We thank Christina Smolke for use of the Molecular Imager and Mona Shahgholi for help with mass spectrometry. Pin Wang and A. James Link kindly provided the purified synthetases. Edman degradation was performed by the Peptide and Protein Molecular Analysis Laboratory of the Beckman Institute at Caltech.

Attached Files

Supplemental Material - z700605_s.pdf

Files

z700605_s.pdf

Files (980.7 kB)

Name	Size	Download all
z700605_s.pdf md5:d4814e4a6dad537a36226750bc9a68d6	980.7 kB	Preview Download

Additional details

	All versions	This version
Views	0	0
Downloads	0	0
Data volume	0 Bytes	0 Bytes