Controlled manipulation of bacteriophages using single-virus force spectroscopy.
A method is described for the site-directed manipulation of single filamentous bacteriophages, by using phage display technology and atomic force microscopy. f1 filamentous bacteriophages were genetically engineered to display His-tags on their pIX tail. Following adsorption on nitrilotriacetate-terminated surfaces, force spectroscopy with tips bearing monoclonal anti-pIII antibodies was used to pull on individual phages via their pIII head. Analysis of the force-extension profiles revealed that upon pulling, the phages are progressively straightened into an extended orientation until rupture of the anti-pIII/pIII complex. The single-virus manipulation technique presented here provides new opportunities for understanding the forces driving cell-virus and material-virus interactions, and for characterizing the binding properties of polypeptide sequences or proteins selected by the phage display technology.
Alsteens
David
D
Unité de Chimie des Interfaces, Université Catholique de Louvain, Croix du Sud 2/18, B-1348 Louvain-la-Neuve, Belgium.
Pesavento
Emanuele
E
Cheuvart
Gilles
G
Dupres
Vincent
V
Trabelsi
Heykel
H
Soumillion
Patrice
P
Dufrêne
Yves F
YF
D000327:Adsorption; D001435:Bacteriophages / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D004926:Escherichia coli / Q000821:virology; D005818:Genetic Engineering; D018625:Microscopy, Atomic Force; D013057:Spectrum Analysis / Q000379:methods*