Modular Protein Ligation: A New Paradigm as a Reagent Platform for Pre-Clinical Drug Discovery

Abstract

Significant resource is spent by drug discovery project teams to generate numerous, yet unique target constructs for the multiple platforms used to drive drug discovery programs including: functional assays, biophysical studies, structural biology, and biochemical high throughput screening campaigns. To improve this process, we developed Modular Protein Ligation (MPL), a combinatorial reagent platform utilizing Expressed Protein Ligation to site-specifically label proteins at the C-terminus with a variety of cysteine-lysine dipeptide conjugates. Historically, such proteins have been chemically labeled non-specifically through surface amino acids. To demonstrate the feasibility of this approach, we first applied MPL to proteins of varying size in different target classes using different recombinant protein expression systems, which were then evaluated in several different downstream assays. A key advantage to the implementation of this paradigm is that one construct can generate multiple final products, significantly streamlining the reagent generation for multiple early drug discovery project teams.

An In Silico Methodology That Facilitates Decision Making in the Engineering of Nanoscale Protein Materials

“…The acquisition and grouping of desired biological activities, which can be combined in a single polypeptide chain through multidomain recruitment, allows producing modular proteins with specific functionalities that are not occurring in nature [1][2][3][4][5][6]. Designing multidomain proteins is a strategy especially appealing in the generation of protein materials at micro-and nanoscales, in which the building blocks must gain assembling abilities for regulatable oligomerization [7,8].…”