Folding and Engineering of Stable and Superstable Proteins

The accurate design and prediction of protein stability are crucial to making useful proteins for biotechnological and medical applications; in addition, a fundamental understanding of protein stability is essential for understanding a multitude of diseases caused by protein misfolding. Our group has been exploring the molecular determinants of stability for ThreeFoil, a 3-fold symmetric, multivalent carbohydrate binding protein which we rationally designed based using consensus, homology, and Rosetta approaches. We find that ThreeFoil has extremely high stability against proteolytic degradation and denaturation by heat, chemicals, and time. Modelling can account for the stability of ThreeFoil and many other proteins. We develop a computational meta-predictor tool to further stabilize ThreeFoil, identify pervasive problems for engineering stability, and suggest how these problems may be solved.