This proposal aims to minimize the three bottlenecks of membrane proteins structure determination with a novel approach that combines high throughput electron microscopy single particle analysis with a novel self-assembling method. Validation of this approach will be realized by application to the structural analysis of several proteins which represent major categories of eukaryotic membrane proteins of biomedical importance. The overall objective of this proposal will be determining their structures and to simultaneously fully develop this approach to tackle further membrane protein targets in our own and other laboratories. The majority of known human pharmaceutical targets are membrane proteins and the future success of structure based drug design efforts will rely heavily on membrane protein structural information. This proposal will minimize the three bottlenecks of membrane proteins structure determination with a novel approach that combines high throughput electron microscopy single particle analysis with a novel self-assembling method. The success of this project will lower the barrier for obtaining the needed information for structure based drug design to proceed for a wide range of diseases.