Application of in Vitro Virus (IVV) Technique for High-Throughput Analysis of Protein-Protein Interactions
Etsuko Miyamoto-Sato and Hiroshi Yanagawa
Global analysis of protein functions and networks has become the focus of considerable attention since the sequencing of the human genome. The development of proteomics has led to an increasing interest in cell-free translation systems because of their rapidity and ease of handling. We have developed the so-called in vitro virus (IVV) as a more stable and efficient tool for evolutionary protein engineering. This system is applicable as an mRNA display technique to analyze protein functions and networks in proteomics. Accordingly, we developed a high-throughput IVV system for analysis of protein-protein interactions employing cell-free co-translation and selection. Here, we overview this system and discuss the advantages of analyzing protein-protein interactions using IVV as a genotype-phenotype assignment molecule in combination with cell-free co-translation. Moreover, we discuss how we address false positives and negatives in this system, and the in silico mass data processing required to derive biologically significant interactions from the wealth of raw data.