Structural studies of the SARS virus Nsp15 endonuclease and the human innate immunity receptor TLR3

• Main Author: Sun, Jingchuan
• Other Authors: Holzenburg, Andreas
• Format: Others
• Language: en_US
• Published: Texas A&M University 2006
• Subjects: Electron Microscopy; 3D reconstruction; 2D crystallization
• Online Access: http://hdl.handle.net/1969.1/3811

Three-dimensional (3D) structural determination of biological macromolecules is not only critical to understanding their mechanisms, but also has practical applications. Combining the high resolution imaging of transmission electron microscopy (TEM) and efficient computer processing, protein structures in solution or in two-dimensional (2D) crystals can be determined. The lipid monolayer technique uses the high affinity binding of 6His-tagged proteins to a Ni-nitrilotriacetic (NTA) lipid to create high local protein concentrations, which facilitates 2D crystal formation. In this study, several proteins have been crystallized using this technique, including the SARS virus Nsp15 endonuclease and the human Toll-like receptor (TLR) 3 extracellular domain (ECD). Single particle analysis can determine protein structures in solution without the need for crystals. 3D structures of several protein complexes had been solved by the single particle method, including IniA from Mycobacterium tuberculosis, Nsp15 and TLR3 ECD. Determining the structures of these proteins is an important step toward understanding pathogenic microbes and our immune system.