{Reference Type}: Journal Article
{Title}: Reconstituted Cell-free Translation Systems for Exploring Protein Folding and Aggregation.
{Author}: Taguchi H;Niwa T;
{Journal}: J Mol Biol
{Volume}: 436
{Issue}: 19
{Year}: 2024 Oct 1
{Factor}: 6.151
{DOI}: 10.1016/j.jmb.2024.168726
{Abstract}: Protein folding is crucial for achieving functional three-dimensional structures. However, the process is often hampered by aggregate formation, necessitating the presence of chaperones and quality control systems within the cell to maintain protein homeostasis. Despite a long history of folding studies involving the denaturation and subsequent refolding of translation-completed purified proteins, numerous facets of cotranslational folding, wherein nascent polypeptides are synthesized by ribosomes and folded during translation, remain unexplored. Cell-free protein synthesis (CFPS) systems are invaluable tools for studying cotranslational folding, offering a platform not only for elucidating mechanisms but also for large-scale analyses to identify aggregation-prone proteins. This review provides an overview of the extensive use of CFPS in folding studies to date. In particular, we discuss a comprehensive aggregation formation assay of thousands of Escherichia coli proteins conducted under chaperone-free conditions using a reconstituted translation system, along with its derived studies.