Advisor
Current Research
Laser-Induced Rehydration of Cryo-Landed Proteins Restores Native Structure
Cryo-electron microscopy (cryo-EM) has become a widely used tool for solving protein structures, but current limitations in sample preparation makes structural analysis impossible for many proteins of interest. Using mass spectrometry (MS) as a sample preparation device, we deposit gas phase proteins onto a cryogenically cooled grid for subsequent analysis with EM. Doing this solves the orientation bias and airwater interface problems present with the commonly used plunge freezing method. However, new challenges present themselves as gas phase proteins undergo structural changes limiting the biological relevance of structures solved with this method. To overcome these imposed challenges, we use a 532nm laser to rapidly melt and revitrify ice deposited on the cryo-EM grid to restore native protein structure and have demonstrated its effectiveness with a model protein. This improvement paves a path forward for MS to be a valuable sample preparation tool for the structural biology community.