A phage display technique for a fast, sensitive, and systematic investigation of protein-protein interactions

作者：Rossenu S; Dewitte D; Vandekerckhove J; Ampe C

来源：Journal of Protein Chemistry, 1997, 16(5): 499-503.

DOI：10.1023/A:1026317612554

• actin
• PCR-mutagenesis
• phage display
• protein-protein interaction
• thymosin beta 4

Phage display is a technique in which a foreign protein or peptide is presented at the surface of a (filamentous) bacteriophage. This system, developed by Smith [(1985), Science 228, 1315-1317], was originally used to create large libraries of antibodies for the purpose of selecting those that strongly bound a particular antigen. More recently it was also employed to present peptides, domains of proteins, or intact proteins at the surface of phages, again to identify high-affinity interactions with ligands. Here we want to illustrate the use of phage display, in combination with PCR saturation mutagenesis, for the study of protein-protein interactions. Rather than selecting for mutants having high affinity, we systematically investigate the binding of every variant with its natural ligand. Via a modified ELISA we can calculate a relative affinity. As a model system we chose to display thymosin beta 4 on the phage surface in order to study its interaction with actin.

• 出版日期1997-7