Knowledge Bank

Most natural proteins are only marginally stable, yet many therapeutic and industrial applications require stabilized variants. It has been shown that mutations to the most common residue in a multiple sequence alignment of protein families results in stabilization about half the time. The degree of conservation of these residues can be determined by measuring the difference between the amino acid distribution at a particular site and its neutral reference state. In this project, we examine if the degree of conservation for a position is related to its ability to affect the stability of the protein. Here, we are interested in studying the additive effects of mutating six positions in yeast triosephosphate isomerase (TIM) to consensus residues. To dissect the effects of these mutations, we are also constructing the six individual mutants which contain one highly conserved residue each. These variants will be studied for both structure and function. With this knowledge we will be able to reliably design more stable and functional proteins from genomic sequence data. This will have numerous benefits for research and industrial purposes, and will have potential health benefits in designing therapeutic proteins.