M13 Bacteriophage Display Framework That Allows Sortase-Mediated Modification of Surface-Accessible Phage Proteins
- 3 July 2012
- journal article
- research article
- Published by American Chemical Society (ACS) in Bioconjugate Chemistry
- Vol. 23 (7), 1478-1487
- https://doi.org/10.1021/bc300130z
Abstract
We exploit bacterial sortases to attach a variety of moieties to the capsid proteins of M13 bacteriophage. We show that pIII, pIX, and pVIII can be functionalized with entities ranging from small molecules (e.g., fluorophores, biotin) to correctly folded proteins (e.g., GFP, antibodies, streptavidin) in a site-specific manner, and with yields that surpass those of any reported using phage display technology. A case in point is modification of pVIII. While a phage vector limits the size of the insert into pVIII to a few amino acids, a phagemid system limits the number of copies actually displayed at the surface of M13. Using sortase-based reactions, a 100-fold increase in the efficiency of display of GFP onto pVIII is achieved. Taking advantage of orthogonal sortases, we can simultaneously target two distinct capsid proteins in the same phage particle and maintain excellent specificity of labeling. As demonstrated in this work, this is a simple and effective method for creating a variety of structures, thus expanding the use of M13 for materials science applications and as a biological tool.Keywords
This publication has 55 references indexed in Scilit:
- Identification of host cell factors required for intoxication through use of modified cholera toxinThe Journal of cell biology, 2011
- Design and Screening of M13 Phage Display cDNA LibrariesMolecules, 2011
- Sortase-catalyzed transformations that improve the properties of cytokinesProceedings of the National Academy of Sciences, 2011
- Site-Specific N- and C-Terminal Labeling of a Single Polypeptide Using Sortases of Different SpecificityJournal of the American Chemical Society, 2009
- Crystal Structure of Streptococcus pyogenes Sortase AJournal of Biological Chemistry, 2009
- Lipid Modification of Proteins through Sortase-Catalyzed TranspeptidationJournal of the American Chemical Society, 2008
- Detection of Early Prostate Cancer Using a Hepsin-Targeted Imaging AgentCancer Research, 2008
- In Vivo Imaging of Molecularly Targeted PhageNeoplasia, 2006
- High copy display of large proteins on phage for functional selections 1 1Edited by P. E. WrightJournal of Molecular Biology, 2000
- Modifying Filamentous Phage Capsid: Limits in the Size of the Major Capsid ProteinJournal of Molecular Biology, 1995