A novel helper phage for HaloTag-mediated co-display of enzyme and substrate on phage

Biochemical and Biophysical Research Communications
Wouter DelespaulJohan Robben

Abstract

Phage display is an established technique for the molecular evolution of peptides and proteins. For the selection of enzymes based on catalytic activity however, simultaneous coupling of an enzyme and its substrate to the phage surface is required. To facilitate this process of co-display, we developed a new helper phage displaying HaloTag, a modified haloalkane dehalogenase that binds specifically and covalently to functionalized haloalkane ligands. The display of functional HaloTag was demonstrated by capture on streptavidin-coated magnetic beads, after coupling a biotinylated haloalkane ligand, or after on-phage extension of a DNA oligonucleotide primer with a biotinylated nucleotide by phi29 DNA polymerase. We also achieved co-display of HaloTag and phi29 DNA polymerase, thereby opening perspectives for the molecular evolution of this enzyme (and others) towards new substrate specificities.

References

May 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·A S KangR A Lerner
Oct 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·L Blanco, M Salas
Jul 23, 1996·Proceedings of the National Academy of Sciences of the United States of America·I FischG Winter
Sep 2, 1998·Proceedings of the National Academy of Sciences of the United States of America·H PedersenP G Schultz
Aug 18, 1999·Proceedings of the National Academy of Sciences of the United States of America·S Atwell, J A Wells
Mar 29, 2001·Proceedings of the National Academy of Sciences of the United States of America·F J GhadessyP Holliger
Apr 18, 2002·Proceedings of the National Academy of Sciences of the United States of America·Frank B DeanRoger S Lasken
May 16, 2002·Proceedings of the National Academy of Sciences of the United States of America·Gang XiaFloyd E Romesberg
Jan 29, 2005·Chembiochem : a European Journal of Chemical Biology·Satoshi FujitaKazunari Taira
Dec 13, 2005·Chembiochem : a European Journal of Chemical Biology·Melloney J DrögeWim J Quax
Mar 22, 2006·Chembiochem : a European Journal of Chemical Biology·Kerry Routenberg LoveSuzanne Walker
Jun 7, 2008·ACS Chemical Biology·Georgyi V LosKeith V Wood
Sep 18, 2009·PloS One·Mark WelchClaes Gustafsson
Jul 16, 2010·Angewandte Chemie·Aaron M LeconteFloyd E Romesberg
Feb 4, 2011·Chembiochem : a European Journal of Chemical Biology·Murat SunbulJun Yin
Mar 27, 2012·Nature Biotechnology·Elizabeth A ManraoJens H Gundlach
Apr 21, 2012·Science·Vitor B PinheiroPhilipp Holliger
Jul 10, 2012·Artificial DNA, PNA & XNA·Yuichiro AibaMakoto Komiyama

❮ Previous
Next ❯

Citations

Dec 21, 2016·Molecular Nutrition & Food Research·Xueni Chen, Stephen C Dreskin
Aug 17, 2016·Biochemical Society Transactions·Pedro A G TizeiVitor B Pinheiro
Mar 5, 2021·Bioconjugate Chemistry·Caitlin M CarmodySam R Nugen

❮ Previous
Next ❯

Related Concepts

Related Feeds

Bacteriophage: Phage Therapy

Phage therapy uses bacterial viruses (bacteriophages) to treat bacterial infections and is widely being recognized as an alternative to antibiotics. Here is the latest research.