Structure and stability of the designer protein WRAP-T and its permutants

Abstract

Abstract $$\beta $$ β -Propeller proteins are common natural disc-like pseudo-symmetric proteins that contain multiple repeats (‘blades’) each consisting of a 4-stranded anti-parallel $$\beta $$ β -sheet. So far, 4- to 12-bladed $$\beta $$ β -propellers have been discovered in nature showing large functional and sequential variation. Using computational design approaches, we created perfectly symmetric $$\beta $$ β -propellers out of natural pseudo-symmetric templates. These proteins are useful tools to study protein evolution of this very diverse fold. While the 7-bladed architecture is the most common, no symmetric 7-bladed monomer has been created and characterized so far. Here we describe such a engineered protein, based on a highly symmetric natural template, and test the effects of circular permutation on its stability. Geometrical analysis of this protein and other artificial symmetrical proteins reveals no systematic constraint that could be used to help in engineering of this fold, and suggests sequence constraints unique to each $$\beta $$ β -propeller sub-family.