Engineering the T1 Domain for Precise, Controlled Assembly of Multidomain Molecular Machines
Our research has identified the T1 domain as the dominant regulator of voltage-gated potassium channel assembly. The T1 domain allows Kv channel alpha subunits within a specific family group to co-assemble promiscuously, while excluding assembly of alpha subunits from another family. While this assembly process provides some degree of control over the final channel complex, the T1 domain does not appear to exert much control on subunit mixing with a family group. By careful engineering of the T1 domain we believe that we can develop an assembly platform that will allow us to construct novel protein machines with precise control over component localization. Using these engineered T1 domains, we will construct novel voltage gated potassium channels with precise control over subunit composition and arrangement. Furthermore, because of the modular nature of voltage gated potassium channels, we believe that these novel T1 domain assembly platforms can be readily transferred to other proteins, allowing the construction of other complex protein machines with precise stereospecific control.
Aim 1: Engineering of T1 domains that assemble with a specified pattern
Aim 2: Engineering of T1 domains with novel interaction properties
Aim 3: Use of Engineered T1 domains to construct heteromultimeric Kv channels with precise control over assembly Aim 4: Exploring ways to make the assembly of Engineered T1 domain machines conditional.
Relevance of the project to IDDRC mission:
The goal of this project is to develop the T1 domain into a flexible nanotechnology assembly platform. Molecular machines developed on this platform should have the ability to be conditionally assembled in neurons and direct specific molecular repair and/or functional changes in situ with precise control. Using such molecular machines, we envision the creation of new molecular machines that can be built within cells to treat the underlying causes of a neuronal defect that leads to an overt phenotype, such as mental retardation.