Precise assembly of complex beta sheet topologies from de novo designed building blocks.

A protein is made up of a sequence of amino acids and must fold into a specific three-dimensional structure if it is to work correctly. The structure is formed by segments of the protein adopting specific shapes, the two most common shapes being alpha helices and beta strands. Beta strands commonly interact with each other to form regions called beta sheets. Researchers trying to design proteins with new abilities have managed to create proteins that contain up to five beta strands and four alpha helices. Larger and more complex proteins are more challenging to make because there are many different ways that a protein can fold. It is also difficult to understand how complex structures such as large beta sheets emerged naturally, over the course of evolution. King et al. have now used computer modeling to explore how a large, complex beta sheet might form. In the model, one small, newly designed protein was inserted into another so that their beta sheets merged to form a single extended sheet. The model then stabilized this structure by changing the amino acids found at the points where the two proteins met. King et al. were then able to synthesize these new proteins in bacteria and use a technique called X-ray crystallography to determine the structure of two of them. The structures closely matched the computer models; one protein contained a six-stranded beta sheet, and the other had a seven-stranded beta sheet. The folds of the two designed proteins were then compared with those found in a database that classifies proteins on the basis of their structure. The beta sheets in the designed proteins did not match the protein structures in the database, which suggests that the designed proteins contained new types of folds. In the future, the technique used by King et al. could be used to design other large and complex beta sheet structures. Furthermore, the results suggest that such large structures could have evolved naturally through the combination of smaller, less complex proteins.