title: M13 bacteriophage display framework that allows sortase-mediated modification of surface-accessible phage proteins.
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.
authors: The David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors: Gaelen T
authors: GT
authors: Hess
authors: Juan J
authors: JJ
authors: Cragnolini
authors: Maximilian W
authors: MW
authors: Popp
authors: Mark A
authors: MA
authors: Allen
authors: Stephanie K
authors: SK
authors: Dougan
authors: Eric
authors: E
authors: Spooner
authors: Hidde L
authors: HL
authors: Ploegh
authors: Angela M
authors: AM
authors: Belcher
authors: Carla P
authors: CP
authors: Guimaraes
mesh_terms: D019881:Aminoacyltransferases / Q000378:metabolism*; D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D017104:Bacteriophage M13 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D061505:Cell Surface Display Techniques / Q000379:methods*; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D013499:Surface Properties
chemical_list: D001426:Bacterial Proteins; D036022:Capsid Proteins; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases