Gregory Winter applied phage display to engineering humanized antibodies and optimizing their therapeutic efficacy ( McCafferty et al., 1990 Clackson et al., 1991 Winter et al., 1994). Gregory Smith developed a phage display method for sorting through millions of peptides or proteins for desired molecular recognition with a target molecule through a process known as biopanning ( Smith, 1985 Smith and Petrenko, 1997). She also engineered cytochrome P450 to catalyze challenging organic reactions ( Kan et al., 2016, 2017 Hammer et al., 2017 Chen et al., 2018). Frances Arnold engineered enzymes by directed evolution to gain tolerance to high temperature or high concentration of organic solvent ( Chen and Arnold, 1993 Zhao et al., 1998).

Winter for fundamental contributions to enzyme-directed evolution and protein engineering. The 2018 Nobel Prize in Chemistry was awarded to Frances H. In this study, we highlight recent achievements of protein engineering on mapping, probing, and manipulating UB transfer in the cell. The recent flourish of protein-engineering approaches such as unnatural amino acid incorporation, protein semisynthesis by expressed protein ligation, and high throughput selection by phage and yeast cell surface display has generated designer proteins as powerful tools to interrogate cell signaling mediated by protein ubiquitination. A greater understanding of the mechanism of UB chain assembly and the signals encoded in UB chains should aid in our understanding of disease pathogenesis and guide the development of novel therapeutics. Dysregulation of the UB transfer pathways and malfunctions of DUBs and UBDs play causative roles in the development of many diseases. Additionally, many cellular proteins carry UB-binding domains (UBDs) that translate the signals encoded in UB chains to target proteins for degradation by proteasomes or in autophagosomes, as well as affect nonproteolytic outcomes such as kinase activation, DNA repair, and transcriptional regulation. Deubiquitinating enzymes (DUBs) reverse UB modifications or trim UB chains of diverse linkages. Ubiquitin (UB) transfer cascades consisting of E1, E2, and E3 enzymes constitute a complex network that regulates a myriad of biologic processes by modifying protein substrates.

Engineered protein scaffold inhibitor