EXPRESSION OF DE-NOVO DESIGNED ALPHA-HELICAL BUNDLES

The successful design of proteins requires careful consideration of th e multiplicity of forces that stabilize their three-dimensional struct ures including hydrophobic interactions, hydrogen-bonding, electrostat ics and weakly polar interactions. Early attempts to design proteins r elied too heavily on hydrophobic interactions to provide stability, re sulting in structures with dynamic properties. Addition of more specif ic interactions to these initial designs gives rise to proteins with m ore native-like properties. This manuscript describes the design of na tive-like three- and four-helix bundles, and their cloning and express ion of these proteins.